Selected renal cell populations, characteristics and uses thereof

Abstract

Methods for identifying enriched heterogeneous renal cell populations with therapeutic potential, enriched heterogeneous renal cell populations with therapeutic potential, and uses thereof.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 512,541, filed July 7, 2023, and U.S. Provisional Patent Application Serial No. 63 / 589,934, filed October 12, 2023. The disclosure of the foregoing reference applications, in its entirety, including any accompanying drawings, is expressly incorporated herein by reference. Background Technology

[0003] Chronic kidney disease (CKD) is on or has reached pandemic scale (Thurlow JS, et al., Am J Nephrol 52(2021):98-107; Murphy D, et al., Intern Med 65(2016):473-481; Hill NR, et al., PLoS One 11:e0158765. doi: 10.1371 / journal.pone.0158765). Prevalence data from the United States to Europe show that approximately 10% of the general population has stage 1-3 CKD (ERA, 2009; USRDS, 2011; Jha et al., Lancet. 382(2013):260-72). In the United States alone, CKD increased by >33% between 1996 and 2006 (US Renal Data System. Costs of CKD and ESRD. Minneapolis, MN, 2007). Meanwhile, the management of CKD remains a challenge for nephrologists. Most standard care treatments for CKD target small molecules in the kidney's biochemical pathways to affect single or related comorbidities. However, these treatments do not address the underlying glomerular and tubulointerstitial dysfunction in the CKD kidney. Ultimately, in patients who progress to end-stage renal disease (ESRD), survival requires renal replacement therapy (RRT; dialysis or transplantation). There remains an unmet need for treatments that directly address the tissue biology of the diseased kidney. Such treatments could potentially halt or even reverse CKD progression and reduce or eliminate the need for RRT.

[0004] Selected renal cells (SRCs), a heterogeneous population of renal cells rich in renal epithelial cells, are being advanced as an autologous cell-based therapy for the treatment of CKD (Stavas J, et al., AJ Nephrol 53(2022):50-58.; Stavas J, et al., KI Reports, in progress). Derived from donor kidneys, SRCs are a mixture primarily composed of proximal tubular epithelial cells (Presnell SC, et al., Tissue Eng Part C Methods 17(2011):261-73). Results from a phase II clinical trial in a diabetic nephropathy cohort indicate that SRC administration is safe and associated with improved renal function (Stavas J, et al., KI Reports, in progress).

[0005] There is a need in the art to identify heterogeneous populations of kidney cells, such as SRCs, that have potential therapeutic activity (e.g., based on their ability to drive tubule formation). Summary of the Invention

[0006] This disclosure generally relates to a method for identifying enriched heterogeneous renal cell populations as having therapeutic potential. In this method, it is determined whether the cells of the enriched heterogeneous renal cell population express at least one cell adhesion marker. The at least one cell adhesion marker includes, or one or more of, the following: intercellular adhesion molecule 5 (ICAM5), melanoma cell adhesion molecule (MCAM), contactin-associated protein 1 (CNTNAP1), or matrix metallopeptidase 2 (MMP2). If the enriched heterogeneous renal cell population is found to express at least one cell adhesion marker, the enriched heterogeneous renal cell population is identified as having therapeutic potential.

[0007] This disclosure describes another method for identifying enriched heterogeneous kidney cell populations as having therapeutic potential. In this method, the expression of transforming growth factor β2 (TGFβ2) in the cells of the enriched heterogeneous kidney cell population is determined. The enriched heterogeneous kidney cell population is identified as having therapeutic potential if it is determined that: (i) more than about 50% of the cells in the enriched heterogeneous kidney cell population express TGFβ2; and / or (ii) TGFβ2 is secreted by the cells of the enriched heterogeneous kidney cell population at a rate of at least about 1.0 ng / 1,000,000 cells in the enriched heterogeneous kidney cell population. Attached Figure Description

[0008] Figure 1Uniform manifold approximation and projection (UMAP) shows the SRC population of cells expressing cdh1. Pink / dark gray dots represent cells expressing cdh1. Light gray dots represent cells not expressing cdh1.

[0009] Figure 2 The image shows the interaction set formed by cell-cell adhesion markers (CDH1, CNTNAP1, PECAM1, ICAM5, MCAM, COL11A1, and MMP2) expressed by human SRC, which is derived from the gene ontology and visualized using miRNet.

[0010] Figure 3 It provides gene ontology mapping, confirming that hsa-miR-145 and hsa-miR-199a-5p regulate tgfβ2 in SRC and renal cell epithelial markers (CDH1, SLC38A2, SLC8A1).

[0011] Figure 4 UMAP shows the SRC cell population expressing tgfβ2. Orange / dark gray dots represent cells expressing tgfβ2. Light gray dots represent cells not expressing tgfβ2.

[0012] Figure 5A and 5B This indicates that TGFβ2 is secreted by SRC. Figure 5A This shows the level or amount of TGFβ2 secreted by human SRC in the culture medium. Figure 5B The study showed that the supernatant from human SRC samples exhibited a time-dependent increase in secreted TGFβ2. p<0.05 relative to 0 hr, #, p<0.05 relative to 72 hr, (p<0.01 relative to 0 hr).

[0013] Figure 6 The gene ontology map reveals that tgfβ2 forms an interaction group with markers of ureteral buds (ret and fgf8) and cap mesenchyme (six2, osr1 and lhx1) expressed by human SRC.

[0014] Figures 7A-7C : This shows that SRCs assemble into small tubes in the culture medium ( Figure 7A ), and assembled small tubes to GGT1 ( Figure 7B ) and SLC12A ( Figure 7C The expression staining was positive.

[0015] Figure 8 The results show the detection of SRC ICAM-1 surface expression by flow cytometry.

[0016] Figure 9 The results show the detection of SRC MCAM surface expression by flow cytometry.

[0017] Figure 10 This indicates that SRC has low E-cadherin surface expression. Invention Details

[0019] This disclosure generally relates to methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, compositions comprising enriched heterogeneous renal cell populations identified as having therapeutic potential, and methods and uses of enriched heterogeneous renal cell populations with therapeutic potential.

[0020] In methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, the therapeutic potential of the enriched heterogeneous renal cell populations can be in the treatment of kidney diseases, tubular transport defects, or glomerular filtration defects.

[0021] The therapeutic potential of the enriched heterogeneous renal cell populations identified by the method may include renal function recovery, stabilization, improvement, reduction of renal fibrosis, reduction of renal inflammation, induction of tubulogenesis, induction of nephrodystrophy, or induction of glomerulogenesis in patients requiring such treatment. The therapeutic potential of the enriched heterogeneous renal cell populations identified by the method may also include the restoration of mineral balance, electrolyte balance, fluid homeostasis, reabsorption of essential nutrients, metabolism of cystatin C, or relief of anemia in patients requiring such treatment. Furthermore, the therapeutic potential of the enriched heterogeneous renal cell populations identified by the method may include the potential to delay or prevent the need for dialysis in patients requiring treatment for kidney disease, or the potential to delay or prevent the need for kidney transplantation.

[0022] In some methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, it is possible to determine whether the cells in the enriched heterogeneous renal cell population express at least one cell adhesion marker. The at least one cell adhesion marker may be, or include any one or more of, the following: intercellular adhesion molecule 5 (ICAM5), melanoma cell adhesion molecule (MCAM), contactin-associated protein 1 (CNTNAP1), or matrix metallopeptidase 2 (MMP2). The at least one cell adhesion marker may be, or may include any one, any two, any three, or all four of the following: ICAM5, MCAM, CNTNAP1, and MMP2.

[0023] In a method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, it is possible to determine whether the cells in the enriched heterogeneous renal cell population express at least two of the cell adhesion markers ICAM5, MCAM, CNTNAP1, and MMP2. In a method for determining the expression of at least two of the cell adhesion markers, the two cell adhesion markers may be or include (a) ICAM5 and MCAM, (b) ICAM5 and CNTNAP1, (c) ICAM5 and MMP2, (d) MCAM and CNTNAP1, (e) MCAM and MMP2, or (f) CNTNAP1 and MMP2.

[0024] In methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, it is possible to determine whether the cells in the enriched heterogeneous renal cell population express at least three of the cell adhesion markers ICAM5, MCAM, CNTNAP1, and MMP2. In methods for determining the expression of at least three of the cell adhesion markers, the three cell adhesion markers may be or include (a) ICAM5, MCAM, and CNTNAP1, (b) ICAM5, MCAM, and MMP2, (c) ICAM5, CNTNAP1, and MMP2, or (d) MCAM, CNTNAP1, and MMP2.

[0025] In methods for identifying enriched heterogeneous kidney cell populations as having therapeutic potential, it is possible to determine whether the cells in the enriched heterogeneous kidney cell populations express cell adhesion markers ICAM5, MCAM, CNTNAP1, and MMP2.

[0026] In a method for identifying enriched heterogeneous kidney cell populations as having therapeutic potential, determining the expression of at least one (e.g., at least one, at least two, at least three, or four) cell adhesion markers in the heterogeneous enriched kidney cell populations can identify them as having therapeutic potential.

[0027] In methods for identifying enriched heterogeneous kidney cell populations as having therapeutic potential, determining the expression of at least one (at least two, at least three, or four) cell adhesion markers may further include determining the percentage of cells in the enriched heterogeneous kidney cell population expressing at least one (at least two, at least three, or four) cell adhesion markers. If the percentage of cells in the enriched heterogeneous kidney cell population expressing at least one (at least two, at least three, or four) cell adhesion markers is determined, then if approximately a certain or specific percentage of the cells in the enriched heterogeneous kidney cell population express at least one (at least two, at least three, or four) cell adhesion markers, the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0028] In this method, if the percentage of cells expressing ICAM5 in an enriched heterogeneous kidney cell population is measured, then if the percentage of cells expressing ICAM5 in the enriched heterogeneous kidney population is greater than 0% and at most about 5%, greater than 0% and at most about 4.5%, greater than 0% and at most about 4%, greater than 0% and at most about 3.5%, greater than 0% and at most about 3%, greater than 0% and at most about 2.5%, greater than 0% and at most about 2%, greater than 0% and at most about 1.8%, greater than 0% and at most about 1%, then the percentage of cells expressing ICAM5 in the enriched heterogeneous kidney population is greater than 0% and at most about 5%, greater than 0% and at most about 4.5%, greater than 0% and at most about 4%, greater than 0% and at most about 3%, greater than 0% and at most about 3%, greater than 0% and at most about 2%, Cells expressing ICAM5 at 0.6%, greater than 0% and at most about 1.4%, greater than 0% and at most about 1.2%, greater than 0% and at most about 1.0%, greater than 0% and at most about 0.8%, greater than 0% and at most about 0.6%, greater than 0% and at most about 0.5%, greater than 0% and at most about 0.4%, greater than 0% and at most about 0.2%, or greater than 0% and at most about 0.1% can identify enriched heterogeneous kidney cell populations as having therapeutic potential.

[0029] The term “about” is used herein to provide literal support for the exact number that follows it, as well as numbers that are close to or approximate the number following the term. In determining whether a number is close to or approximates a specifically stated number, a number that is close to or approximates an unstated number may be a number provided in the context in which it is presented that is substantially equivalent to the specifically stated number. If the degree of approximation is unclear from the context, “about” means within 10% of the provided value, or rounded to the nearest significant figure, in all cases including the provided value. In some embodiments, the term “about” means a specified value ± at most 10%, at most ± 5%, or at most ± 1%.

[0030] In this method, if the percentage of cells expressing ICAM5 in an enriched heterogeneous kidney cell population is measured, then if more than about 50%, such as more than about 55%, more than about 60%, more than about 65%, more than about 70%, more than about 75%, more than about 80%, more than about 85%, more than about 90%, more than about 95%, more than about 96%, more than about 97%, more than about 98%, or more than about 99% of the cells in the enriched heterogeneous kidney cell population express ICAM5, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0031] In this method, if the percentage of cells expressing MCAM in an enriched heterogeneous kidney cell population is measured, then the percentages are: greater than 10% and at most approximately 35%, greater than approximately 10% and at most approximately 32.5%, greater than approximately 10% and at most approximately 30%, greater than approximately 10% and at most approximately 27.5%, greater than approximately 10% and at most approximately 25%, greater than approximately 12.5% ​​and at most approximately 35%, greater than approximately 12.5% ​​and at most approximately 32.5%, greater than approximately 12.5% ​​and at most approximately 30%, greater than approximately 12.5% ​​and at most approximately 27.5%, and greater than approximately 12.5%. Cells expressing MCAM at most about 25%, greater than about 15% and at most about 35%, greater than about 15% and at most about 32.5%, greater than about 15% and at most about 30%, greater than 15% and at most about 27.5%, greater than about 15% and at most about 25%, greater than about 17.5% and at most about 35%, greater than about 17.5% and at most about 32.5%, greater than about 17.5% and at most about 30%, greater than about 17.5% and at most about 27.5%, or greater than about 17.5% and at most about 25% can identify enriched heterogeneous kidney cell populations as having therapeutic potential.

[0032] In this method, if the percentage of cells expressing MCAM in an enriched heterogeneous kidney cell population is determined, then if more than about 50%, such as more than about 55%, more than about 60%, more than about 65%, more than about 70%, more than about 75%, more than about 80%, more than about 85%, more than about 90%, more than about 95%, more than about 96%, more than about 97%, more than about 98%, or more than about 99% of the cells in the enriched heterogeneous kidney cell population express MCAM, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0033] In this method, if the percentage of cells expressing CNTNAP1 in an enriched heterogeneous kidney cell population is determined, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential if more than about 2% and at most about 20%, more than about 2% and at most about 17.5%, more than about 2% and at most about 15%, more than about 2% and at most about 12.5%, more than about 2% and at most about 10%, more than about 4% and at most about 20%, more than about 4% and at most about 17.5%, more than about 4% and at most about 15%, more than about 4% and at most about 12.5%, more than about 4% and at most about 10%, more than about 6% and at most about 20%, more than about 6% and at most about 17.5%, more than about 6% and at most about 15%, more than about 6% and at most about 12.5%, or more than about 6% and at most about 10% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1.

[0034] In this method, if the percentage of cells expressing CNTNAP1 in an enriched heterogeneous kidney cell population is determined, then if more than about 10% and at most about 40%, more than about 10% and at most about 37.5%, more than about 10% and at most about 35%, more than about 10% and at most about 32.5%, more than about 10% and at most about 30%, more than about 12.5% ​​and at most about 40%, more than about 12.5% ​​and at most about 37.5%, more than about 12.5% ​​and at most about 35%, more than about 12.5% ​​and at most about 32.5%, more than about 15% and at most about 40%, more than about 15% and at most about 34.5%, more than about 15% and at most about 35%, or more than about 15% and at most about 32.5% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0035] In the method, if the percentage of cells expressing MMP2 in the enriched heterogeneous kidney cell population is measured, then if the percentage of cells expressing MMP2 in the enriched heterogeneous kidney cell population is greater than about 5% and at most about 30%, greater than about 5% and at most about 27.5%, greater than about 5% and at most about 25%, greater than about 5% and at most about 22.5%, greater than about 5% and at most about 20%, greater than about 7.5% and at most about 30%, greater than about 7.5% and at most about 27.5%, greater than Cells expressing MMP2 at approximately 7.5% and up to approximately 25%, greater than approximately 7.5% and up to approximately 22.5%, greater than approximately 7.5% and up to approximately 20%, greater than approximately 10% and up to approximately 30%, greater than approximately 10% and up to approximately 27.5%, greater than approximately 10% and up to approximately 25%, greater than approximately 10% and up to approximately 22.5%, or greater than approximately 10% and up to approximately 20% can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0036] In this method, if the percentage of cells expressing MMP2 in an enriched heterogeneous kidney cell population is determined, then if more than about 8% and up to about 40%, more than about 8% and up to about 37.5%, more than about 8% and up to about 35%, more than about 8% and up to about 32.5%, more than about 8% and up to about 30%, more than about 10% and up to about 40%, more than about 10% and up to about 37.5%, more than about 10% and up to about 35%, more than about 10% and up to about 32.5%, more than about 10% and up to about 30%, more than about 12% and up to about 40%, more than about 12% and up to about 37.5%, more than about 12% and up to about 35%, more than about 12% and up to about 32.5%, and more than about 10% and up to about 30% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0037] In this method, if the percentage of cells expressing MMP2 in an enriched heterogeneous kidney cell population is measured, then the percentages of cells expressing MMP2 in the enriched heterogeneous kidney cell population are: greater than approximately 13% and at most approximately 40%, greater than approximately 13% and at most approximately 37.5%, greater than approximately 13% and at most approximately 35%, greater than approximately 13% and at most approximately 32.5%, greater than approximately 13% and at most approximately 30%, greater than approximately 15% and at most approximately 40%, and greater than approximately 15% and at most approximately 37.5%. Cells expressing MMP2 at a rate greater than approximately 15% and up to approximately 35%, greater than approximately 15% and up to approximately 32.5%, greater than approximately 15% and up to approximately 30%, greater than approximately 17% and up to approximately 40%, greater than approximately 17% and up to approximately 37.5%, greater than approximately 17% and up to approximately 35%, greater than approximately 17% and up to approximately 32.5%, or greater than approximately 17% and up to approximately 30% can be identified as enriched heterogeneous renal cell populations with therapeutic potential.

[0038] In the method, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if the percentage of cells expressing at least one (e.g., at least one, or at least two, or at least three, or all four) of the cell adhesion markers ICAM5, MCAM, CNTNAP, and MMP2 in a heterogeneous kidney cell population is determined, and if any combination of any two, any three, or all four is determined: (i) more than 0% and at most about 2% of the cells in the heterogeneous kidney cell population express ICAM5, (ii) more than about 10% and at most about 35% of the cells in the heterogeneous kidney cell population express MCAM, (iii) more than about 2% and at most about 20% of the cells in the heterogeneous kidney cell population express CNTNAP1, and / or (iv) more than about 5% and at most about 30% of the cells in the heterogeneous kidney cell population express MMP2.

[0039] In another instance, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if any two, any three, or a combination of all four of the following are measured: (i) greater than about 0% and at most about 0.5% of cells in the heterogeneous kidney cell population express ICAM5, (ii) greater than about 15% and at most about 30% of cells in the heterogeneous kidney cell population express MCAM, (iii) greater than about 4% and at most about 15% of cells in the heterogeneous kidney cell population express CNTNAP1, and / or (iv) greater than about 7.5% and at most about 25% of cells in the heterogeneous kidney cell population express MMP2. In yet another instance, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if any two, any three, or all four of the following conditions are measured: (i) greater than about 0% and at most about 0.4% of cells in the heterogeneous kidney cell population express ICAM5, (ii) greater than about 17.5% and at most about 27.5% of cells in the heterogeneous kidney cell population express MCAM, (iii) greater than about 4% and at most about 12.5% ​​of cells in the heterogeneous kidney cell population express CNTNAP1, and / or (iv) greater than about 10% and at most about 20% of cells in the heterogeneous kidney cell population express MMP2.

[0040] In yet another instance, if any one, any two, any three, any four, any five, or all six of the following conditions are measured: (a) more than about 60% of the enriched heterogeneous kidney cell population expresses ICAM5 and more than about 65% of the enriched heterogeneous kidney cell population expresses MCAM; (b) more than about 60% of the enriched heterogeneous kidney cell population expresses ICAM5 and more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population expresses CNTNAP1; (c) more than about 60% of the enriched heterogeneous kidney cell population expresses ICAM5 and more than about 10% and at most about 35% of the enriched heterogeneous kidney cell population expresses MCAM. An enriched heterogeneous kidney cell population can be identified as having therapeutic potential if: (d) more than 65% of the cells in the enriched heterogeneous kidney cell population express MCAM and more than 15% and at most 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1; (e) more than 65% of the cells in the enriched heterogeneous kidney cell population express MCAM and more than 10% and at most 35% of the cells in the enriched heterogeneous kidney cell population express MMP2; and / or (f) more than 15% and at most 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1 and more than 10% and at most 35% of the cells in the enriched heterogeneous kidney cell population express MMP2; or (f) more than 15% and at most 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1 and more than 10% and at most 35% of the cells in the enriched heterogeneous kidney cell population express MMP2.

[0041] In yet another instance, if any one, any two, any three, or all four of the following conditions are measured: (a)(i) greater than 0% and at most about 0.5% of cells in an enriched heterogeneous kidney cell population express ICAM5, (ii) greater than about 15% and at most about 30% of cells in an enriched heterogeneous kidney cell population express MCAM, and (iii) greater than about 4% and at most about 15% of cells in an enriched heterogeneous kidney cell population express CNTNAP1; (b)(i) greater than 0% and at most about 0.5% of cells in an enriched heterogeneous kidney cell population express ICAM5, (ii) greater than about 15% and at most about 30% of cells in an enriched heterogeneous kidney cell population express MCAM, and (iii) greater than about 8% and at most about 25% of cells in an enriched heterogeneous kidney cell population express ICAM5. The enriched heterogeneous kidney cell population can be identified as having therapeutic potential if: (c)(i) more than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population expresses ICAM5; (ii) more than about 4% and at most about 15% of the enriched heterogeneous kidney cell population expresses CNTNAP1; and (iii) more than about 8% and at most about 25% of the enriched heterogeneous kidney cell population expresses MMP2; and / or (d)(i) more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population expresses MCAM; (ii) more than about 4% and at most about 15% of the enriched heterogeneous kidney cell population expresses CNTNAP; and (iii) more than about 8% and at most about 25% of the enriched heterogeneous kidney cell population expresses MMP2.

[0042] In yet another instance, if any one, any two, any three, or all four of the following conditions are determined: (a)(i) more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, (ii) more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, and (iii) more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1; (b)(i) more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, (ii) more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, and (iii) more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2; c)(i) greater than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, (ii) greater than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, and (iii) greater than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2; and / or (d)(i) greater than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM; (ii) greater than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP; (iii) greater than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0043] In another instance, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) more than 0% and at most about 0.5% of the cells in the enriched heterogeneous kidney cell population express ICAM5, (ii) more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express MCAM, (iii) more than about 4% and at most about 15% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, and (iv) more than about 8% and at most about 25% of the cells in the enriched heterogeneous kidney cell population express MMP2.

[0044] In another instance, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, (ii) more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, (iii) more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, and (iv) more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2.

[0045] The expression of the markers can be determined in these percentages. Any combination of any two, any three, or all four markers can be a combination of any of the following: (a) ICAM5 and MCAM; (b) ICAM5 and CNTNAP1; (c) ICAM5 and MMP2; (d) MCAM and CNTNAP1; (e) MCAM and MMP2; (f) CNTNAP1 and MMP2; (g) ICAM5, MCAM, and CNTNAP1; (h) ICAM5, MCAM, and MMP2; (i) ICAM5, CNTNAP1, and MMP2; (j) MCAM, CNTNAP1, and MMP2; or (k) ICAM5, MCAM, CNTNAP1, and MMP2.

[0046] In methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, in addition to determining whether the cells of the enriched heterogeneous renal cell population express at least one cell adhesion marker (e.g., any one of the following: ICAM5; or MCAM; or CNTNAP1; or MMP2; or ICAM5 and MCAM; or ICAM5 and CNTNAP1; or ICAM5 and MMP2; or MCAM and CNTNAP1; or MCAM and MMP2; or CNTNAP1 and MMP2; or ICAM5, MCAM and CNTNAP1; or ICAM5, MCAM and MMP2; or ICAM5, CNTNAP1 and MMP2; or MCAM, CNTNAP1 and MMP2; or ICAM5, MCAM, CNTNAP1 and MMP2), it may be further determined whether the cells of the enriched heterogeneous renal cell population express at least one renal-derived marker, wherein at least one renal-derived marker is one or more of the following: sine oculis homolog 2 (sine oculis homolog 2). The markers used include homeobox homolog 2 (SIX2), rearranged during transfection (RET), odd-skipped related 1 (OSR1), lim homeobox protein 1 (LHX1), or fibroblast growth factor 8 (FGF8). Further expression of at least one renal marker in enriched heterogeneous cells can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0047] The method may further measure at least one renal biomarker expressing the biomarker, which may be any one of SIX2, RET, OSR1, LHX1, or FGF8. Alternatively, the method may further measure at least one renal biomarker expressing the biomarker, which may be any one of the following two renal biomarkers or may include any one of the following two renal biomarkers: (a) SIX2 and OSR1; (b) SIX2 and LHX1; (c) SIX2 and RET; (d) SIX2 and FGF8; (e) OSR1 and LHX1; (f) OSR1 and RET; (g) OSR1 and FGF8; (h) LHX1 and RET; (i) LHX1 and FGF8; (j) RET and FGF8. The method may further determine at least one renal biomarker expressing the biomarker, which may be any one of the following three renal biomarkers or may include any one of the following three renal biomarkers: (a) SIX2, OSR1 and LHX1, (b) SIX2, OSR1 and RET, (c) SIX2, OSR1 and FGF8, (d) SIX2, LHX1 and RET, (e) SIX2, LHX1 and FGF8, (f) SIX2, RET and FGF8, (g) OSR1, LHX1 and RET, (h) OSR1, LHX1 and FGF8, (i) OSR1, RET and FGF8, or (j) LHX1, RET and FGF8. The at least one renal biomarker that can be further measured in this method may be any one of the following four renal biomarkers, or may include any one of the following four renal biomarkers: (a) SIX2, OSR1, LHX1, and RET; (b) SIX2, OSR1, LHX1, and FGF8; (c) SIX2, LHX1, RET, and FGF8; (d) SIX2, OSR1, RET, and FGF8; or (e) OSR1, LHX1, RET, and FGF8. The at least one renal biomarker that can be further measured in this method may include five renal biomarkers: SIX2, OSR1, LHX1, RET, and FGF8.

[0048] In a method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, the expression of at least one renal biomarker (at least two, at least three, or four cell adhesion biomarkers in addition to at least one) is further determined. The determination of the expression of at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers may further include determining the percentage of cells in the enriched heterogeneous renal cell population expressing at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers. If the percentage of cells in the enriched heterogeneous renal cell population expressing at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers is further determined, then if approximately a certain or specific percentage of cells in the enriched heterogeneous renal cell population express at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0049] In the case of further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in an enriched heterogeneous renal cell population, if at least one (at least two, at least three, at least four, or five) renal marker is or includes SIX2, then if further determining the percentage of cells expressing at least 0% and at most about 10%, or at least about 0% and at most about 8%, or at least 0% and at most about 6%, or at least 0% and at most about 5.5%, or at least 0% and at most about 5.5%, or at least 0% and at most about 5.5%, then the percentage of cells expressing at least 0% and at most about 10% in the enriched heterogeneous renal cell population is considered to be greater than 0% and at most about 10%, or greater than about 0% and at most about 8%, or greater than 0% and at most about 6%, or greater than 0% and at most about 5.5%, or greater than 0% and at most about 5.5%, then the percentage of cells expressing at least 0% and at most about 5.5% in the enriched heterogeneous renal cell population is considered to be greater than 0% and at most about 10%, or greater than about 0% and at most about 8%, or greater than 0% and at most about 6%, or greater than 0% and at most about 5.5%, then the percentage of cells expressing at least 0% and at most 5.5% in the enriched heterogeneous renal cell population is considered to be greater than 0% and at most about 10%, or greater than about 0% and at most about 5.5%, or greater than 0% and at most about 5.5%, then the percentage of cells expressing at least 0% and at most 5.5% in the enriched heterogeneous renal cell population is considered to be greater than 0% and at most about 5.5%, or greater than 0% and at most about 5.5%, then the percentage of cells expressing Cells expressing SIX2 at approximately 5%, or greater than 0% and at most about 4.5%, or greater than 0% and at most about 4%, or greater than 0% and at most about 3.5%, or greater than 0% and at most about 3%, or greater than 0% and at most about 2.5%, or greater than 0% and at most about 2%, or greater than 0% and at most about 1.5%, or greater than 0% and at most about 1%, or greater than 0% and at most about 0.8%, or greater than 0% and at most about 0.6%, or greater than 0% and at most about 0.4%, or greater than 0% and at most about 0.2% can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0050] In the case of further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population, if at least one (at least two, at least three, at least four, or five) renal marker is or includes OSR1, then if further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population is greater than 0% and at most about 90%, greater than 0% and at most about 85%, greater than 0% and at most about 80%, greater than 0% and at most about 75%, greater than 0% and at most about 70%, greater than 0% and at most about 65%, greater than 0% and at most about 60%, greater than 0% and at most about 55%, greater than 0% and at most about 50%, greater than 0% and at most about 45%, greater than 0% and at most about 40%, greater than 0% and at most about 35%, greater than 0% and at most about 30%, greater than 0% and at most about ... 25%, greater than 0% and at most about 20%, greater than 0% and at most about 15%, greater than 0% and at most about 10%, greater than about 0.15% and at most about 8%, greater than about 0.15% and at most about 9%, greater than about 0.15% and at most about 10%, greater than about 0.15% and at most about 11%, greater than about 0.15% and at most about 12%, greater than about 35% and at most about 90%, greater than about 35% and at most about 85%, greater than about 35% If up to approximately 80%, greater than approximately 35% and up to approximately 75%, greater than approximately 35% and up to approximately 70%, greater than approximately 40% and up to approximately 90%, greater than approximately 45% and up to approximately 90%, greater than approximately 50% and up to approximately 90%, greater than approximately 55% and up to approximately 90%, greater than approximately 60% and up to approximately 90%, or greater than approximately 65% ​​and up to approximately 90% of cells express OSR1, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0051] In the case of further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population, if at least one (at least two, at least three, at least four, or five) renal marker is or includes LHX1, then if further determining the percentage of cells expressing at least 5% and at most about 90%, at most about 5% and at most about 85%, at most about 5% and at most about 80%, at most about 5% and at most about 75%, at most about 5% and at most about 75%, at most about 5% and at most about 90%, at most about 5% and at most about 85%, at most about 5% and at most about 75%, at most about 5% and at most about 90 ... Approximately 5% and at most approximately 70%, greater than approximately 5% and at most approximately 65%, greater than approximately 5% and at most approximately 60%, greater than approximately 5% and at most approximately 55%, greater than approximately 5% and at most approximately 50%, greater than approximately 5% and at most approximately 45%, greater than approximately 5% and at most approximately 40%, greater than approximately 10% and at most approximately 90%, greater than approximately 10% and at most approximately 85%, greater than approximately 10% and at most approximately 80%, greater than approximately 10% and at most approximately 75%, greater than approximately 10% and at most approximately 70%, greater than approximately 10% and at most approximately 65%, greater than approximately 10% and at most approximately 60%, greater than approximately 10% and at most approximately 60%, greater than approximately 10% and Up to approximately 55%, greater than approximately 10% and up to approximately 50%, greater than approximately 10% and up to approximately 45%, greater than approximately 10% and up to approximately 40%, greater than approximately 15% and up to approximately 90%, greater than approximately 15% and up to approximately 85%, greater than approximately 15% and up to approximately 80%, greater than approximately 15% and up to approximately 75%, greater than approximately 15% and up to approximately 70%, greater than approximately 15% and up to approximately 65%, greater than approximately 15% and up to approximately 60%, greater than approximately 15% and up to approximately 55%, greater than approximately 15% and up to approximately 50%, greater than approximately 15% and up to approximately 45%, greater than approximately 15% and up to approximately 45%, greater than approximately 15% and up to approximately 50 ... Cells expressing LHX1 can be identified as having therapeutic potential if they are at least 40% or more, greater than about 20% and at most about 90%, greater than about 20% and at most about 85%, greater than about 20% and at most about 80%, greater than about 20% and at most about 75%, greater than about 20% and at most about 70%, greater than about 20% and at most about 65%, greater than about 20% and at most about 60%, greater than about 20% and at most about 55%, greater than about 20% and at most about 50%, greater than about 20% and at most about 45%, or greater than about 20% and at most about 40%.

[0052] In the case of further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population, if at least one (at least two, at least three, at least four, or five) renal marker is or includes RET, then if further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population is greater than 0% and at most about 90%, greater than 0% and at most about 80%, greater than 0% and at most about 70%, greater than 0% and at most about 60%, greater than 0% and at most about 50%, greater than 0% and at most about 45%, greater than 0% and at most about 40%, greater than 0% and at most about 35%, greater than 0% and at most about 30%, greater than 0% and at most about 25%, greater than 0% and at most about 20%, or greater than 0% and at most about 15%, then the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population is greater than 0% and at most about 15%. Cells expressing RET at a rate greater than 0% and up to about 10%, greater than 0% and up to about 9%, greater than 0% and up to about 8%, greater than 0% and up to about 7%, greater than 0% and up to about 6%, greater than 0% and up to about 5%, greater than 0% and up to about 4%, greater than 0% and up to about 3%, greater than 0% and up to about 2%, greater than 0% and up to about 1%, greater than about 0.2% and up to about 10%, greater than about 0.2% and up to about 9%, greater than about 0.2% and up to about 8%, greater than about 0.2% and up to about 7%, greater than about 0.2% and up to about 6%, greater than about 0.2% and up to about 5%, greater than about 0.2% and up to about 4%, greater than about 0.2% and up to about 3%, greater than about 0.2% and up to about 2%, or greater than about 0.2% and up to about 1% can identify enriched heterogeneous kidney cell populations as having therapeutic potential.

[0053] In the case of further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population, if at least one (at least two, at least three, at least four, or five) renal marker is or includes FGF8, then if further determining the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in the enriched heterogeneous renal cell population is greater than 0% and at most about 60%, greater than 0% and at most about 50%, greater than 0% and at most about 40%, greater than 0% and at most about 30%, greater than 0% and at most about 20%, greater than 0% and at most about 10%, greater than 0% and at most about 9%, greater than 0% and at most about 6 ... Cells expressing FGF8 at approximately 8%, greater than 0% and at most approximately 7%, greater than 0% and at most approximately 6%, greater than 0% and at most approximately 5%, greater than 0% and at most approximately 4%, greater than 0% and at most approximately 3%, greater than 0% and at most approximately 2%, greater than 0% and at most approximately 1%, greater than 0% and at most approximately 0.9%, greater than 0% and at most approximately 0.8%, greater than 0% and at most approximately 0.7%, greater than 0% and at most approximately 0.6%, greater than 0% and at most approximately 0.5%, greater than 0% and at most approximately 0.4%, greater than 0% and at most approximately 0.3%, greater than 0% and at most approximately 0.2%, or greater than 0% and at most approximately 0.1% can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0054] For example, in the case of a method for further determining the percentage of cells expressing at least one (at least one, or at least two, or at least three, or at least four, or five) renal markers in an enriched heterogeneous renal cell population, if any two, any three, any four, or all of the following are further determined: (i) more than 0% and at most about 0.2% of the cells in the enriched heterogeneous renal cell population express SIX2, (ii) more than about 0.015% and at most about 11% of the cells in the enriched heterogeneous renal cell population express OSR1, (iii) more than about 20% and at most about 40% of the cells in the enriched heterogeneous renal cell population express LHX1, (iv) more than about 0.2% and at most about 0.6% of the cells in the enriched heterogeneous renal cell population express RET and / or (v) more than 0% and at most about 0.3% of the cells in the enriched heterogeneous renal cell population express FGF8, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. In another example of a method in which the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal markers in an enriched heterogeneous renal cell population is further determined, the enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all five of the following are further determined: (i) greater than 0% and at most about 6% of the cells in the enriched heterogeneous renal cell population express SIX2; (ii) greater than 0% and at most about 85% of the cells in the enriched heterogeneous renal cell population express OSR1; (iii) greater than about 8% and at most about 90% of the cells in the enriched heterogeneous renal cell population express LHX1; (iv) greater than 0% and at most about 90% of the cells in the enriched heterogeneous renal cell population express RET; and / or (v) greater than 0% and at most about 60% of the cells in the enriched heterogeneous renal cell population express FGF8. In another example of a method in which the percentage of cells expressing at least one (at least two, at least three, at least four, or five) renal biomarkers in an enriched heterogeneous renal cell population is further determined, the enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all five of the following are further determined: (i) more than about 0.1% and at most about 10% of the cells in the enriched heterogeneous renal cell population express SIX2; (ii) more than 0.15% and at most about 80% of the cells in the enriched heterogeneous renal cell population express OSR1; (iii) more than about 8% and at most about 58% of the cells in the enriched heterogeneous renal cell population express LHX1; (iv) more than 10% and at most about 80% of the cells in the enriched heterogeneous renal cell population express RET; and / or (v) more than 0% and at most about 5% of the cells in the enriched heterogeneous renal cell population express FGF8.The expression of these markers can be further determined in these percentages. Any two, three, four, or all five markers can be a combination of any of the following: SIX2 and OSR1; SIX2 and LHX1; SIX2 and RET; SIX2 and FGF8; OSR1 and LHX1; OSR1 and RET; OSR1 and FGF8; LHX1 and RET; LHX1 and FGF8; RET and FGF8; SIX2, OSR1 and LHX1; SIX2, OSR1 and RET; SIX2, OSR1 and FGF8; SIX2, LHX1 and RET ;SIX2, LHX1 and FGF8;SIX2, RET and FGF8;OSR1, LHX1 and RET;OSR1, LHX1 and FGF8;OSR1, RET and FGF8;LHX1, RET and FGF8;SIX2, OSR1, LHX1 and RET;SIX2, OSR1, LHX1 and FGF8;SIX2, LHX1, RET and FGF8;SIX2, OSR1, RET and FGF8;OSR1, LHX1, RET and FGF8;orSIX2, OSR1, LHX1, RET and FGF8.

[0055] In methods for determining the therapeutic potential of enriched heterogeneous renal cell populations, determining the expression of at least one cell adhesion marker (at least two, at least three, or four of ICAM5, MCAM, CNTNAP1, or MMP2) may further determine the expression of at least one additional cell adhesion marker, wherein the at least one additional cell adhesion marker may include any one or more of the following: type XI collagen α1 (COL11A1), platelet endothelial cell adhesion molecule (PECAM) 1, or E-cadherin. Determining the expression of at least one cell adhesion marker and at least one additional cell adhesion marker can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0056] In some cases, determining the expression of at least one additional cell adhesion molecule (in addition to at least one, at least two, at least three, or four of ICAM5, MCAM, CNTNAP1, and / or MMP2) may be done by determining the percentage of cells expressing at least one additional cell adhesion marker in an enriched heterogeneous kidney cell population. In each case, the percentage of cells expressing at least one additional cell adhesion marker in an enriched heterogeneous kidney cell population is determined, and the additional cell adhesion molecule may be or may include COL11A1. In these cases, the percentage of enriched heterogeneous renal cell populations expressing COL11A1 that can be further determined to have therapeutic potential can be greater than about 15% and up to about 40%, greater than about 16% and up to about 40%, greater than about 17% and up to about 40%, greater than about 18% and up to about 40%, greater than about 19% and up to about 40%, greater than about 20% and up to about 40%, greater than about 20% and up to about 41%, greater than about 20% and up to about 42%, greater than about 20% and up to about 43%, greater than about 20% and up to about 44%, greater than about 20% and up to about 45%, greater than 20% and up to about 46%, greater than 20% and up to about 48%, and greater than 20% and up to about 45%. More than 50%, greater than 20% and at most about 52%, greater than 20% and at most about 54%, greater than 20% and at most about 55%, greater than about 25% and at most about 40%, greater than about 25% and at most about 42%, greater than 25% and at most about 44%, greater than about 25% and at most about 46%, greater than about 25% and at most about 48%, greater than about 25% and at most about 50%, greater than about 25% and at most about 52%, greater than about 25% and at most about 54%, greater than about 25% and at most about 55%, greater than about 15% and at most about 41%, greater than about 15% and at most about 42%, greater than about 15% and at most about 43%, greater than about 15% and at most about 44%, greater than about 15% and at most about 45%.

[0057] In cases where the percentage of cells expressing at least one additional cell adhesion marker in an enriched heterogeneous renal cell population is determined and the additional cell adhesion molecule is or includes PECAM1, the percentage of cells expressing PECAM1 that can be further determined to identify the enriched heterogeneous renal cell population as having therapeutic potential can be greater than 0% and up to about 5%, greater than 0% and up to about 4%, greater than 0% and up to about 3%, greater than 0% and up to about 2%, greater than 0% and up to about 1%, greater than 0% and up to about 0.5%, greater than 0% and up to about 0.4%, or greater than 0% and up to about 0.3%.

[0058] In cases where the percentage of cells expressing at least one additional cell adhesion marker in an enriched heterogeneous renal cell population is determined, and the additional cell adhesion molecule is or includes CDH1 (also known as E-cadherin), the percentage of CDH1-expressing cells in the enriched heterogeneous renal cell population can be further determined to identify the percentage of CDH1-expressing cells with therapeutic potential as greater than about 8% and up to about 40%, greater than about 8% and up to about 39%, greater than about 8% and up to about 38%, greater than about 8% and up to about 37%, greater than about 8% and up to about 36%, greater than about 8% and up to about 35%, greater than about 10% and up to about 20%, greater than about 10% and up to about 19%, greater than about 10% and up to about 18%, and greater than about 10% and up to Approximately 17%, greater than approximately 10% and at most approximately 40%, greater than approximately 11% and at most approximately 40%, greater than approximately 12% and at most approximately 40%, greater than approximately 13% and at most approximately 40%, greater than approximately 14% and at most approximately 40%, greater than approximately 15% and at most approximately 40%, greater than approximately 16% and at most approximately 40%, greater than approximately 17% and at most approximately 40%, greater than approximately 18% and at most approximately 40%, greater than approximately 19% and at most approximately 40%, greater than approximately 15% and at most approximately 39%, greater than approximately 15% and at most approximately 38%, greater than approximately 15% and at most approximately 37%, greater than approximately 15% and at most approximately 36%, greater than approximately 15% and at most approximately 35%, greater than approximately 15% and at most approximately 34%, or greater than approximately 15% and at most approximately 33%. In such cases, the percentage of cells expressing CDH1 can be the percentage of cells expressing CDH1 mRNA. In other cases, the percentage of cells expressing CDH1 can be greater than 0% and at most about 5%, or greater than 0% and at most about 4.5%, or greater than about 0% and at most about 4%, greater than 0% and at most about 3.5%, greater than about 0% and at most about 3.0%, greater than 0% and at most about 2.5%, greater than about 0% and at most about 2%, greater than 0% and at most about 1.5%, greater than 0% and at most about 1%, or greater than 0% and at most about 0.5%. In these cases, the percentage of cells expressing CDH1 can be the percentage of cells expressing the CDH1 protein on their surface.

[0059] In methods for determining the therapeutic potential of enriched heterogeneous renal cell populations, the assay may include not only determining the expression of at least one, at least two, at least three, or four cell adhesion markers (any one of ICAM5, MCAM, CNTNAP1, and / or MMP2) and optionally at least one renal-derived marker (any one, at least two, at least three, at least four, or five of SIX1, RET, OSR1, LHX1, and / or FGF8), but also optionally at least one additional cell adhesion marker (any one, two, or three of COL11A1, PECAM1, and / or E-cadherin). Optionally, it may further include determining the expression of one or more additional markers. Examples of further markers include, but are not limited to, TGFβ2, nephrin, podocin, or the receptor for activated C kinase (RACK-1). Further determination of the expression of at least one additional marker may identify enriched heterogeneous renal cell populations as having therapeutic potential. In some cases, further determination of the percentage of cells expressing at least one additional marker in an enriched heterogeneous kidney cell population may be necessary.

[0060] In cases where the percentage of cells expressing at least one additional marker in the enriched heterogeneous renal cell population is or includes TGFβ2, the percentage of TGFβ2-expressing cells in the enriched heterogeneous renal cell population can be identified as having therapeutic potential, and can be greater than about 40% and at most about 85%, greater than about 40% and at most about 80%, greater than about 40% and at most about 75%, greater than about 40% and at most about 70%, or greater than about 45%. And at most about 85%, greater than about 45% and at most about 80%, greater than about 45% and at most about 75%, greater than about 45% and at most about 70%, greater than about 50% and at most about 85%, greater than about 50% and at most about 80%, greater than about 50% and at most about 75%, greater than about 50% and at most about 70%, greater than about 55% and at most about 85%, greater than about 50% and at most about 80%, greater than about 50% and at most about 75%, or greater than about 50% and at most about 80%.

[0061] In cases where the percentage of cells expressing at least one additional marker in an enriched heterogeneous renal cell population is determined, and the additional marker is or includes a method for identifying nephrotic proteins, the percentage of cells expressing nephrotic proteins in the enriched heterogeneous renal cell population that express therapeutic potential can be greater than about 68%, greater than about 70%, greater than about 72%, greater than about 74%, greater than about 76%, greater than about 78%, greater than about 80%, greater than about 82%, greater than about 84%, greater than about 86%, greater than about 88%, greater than about 90%, greater than about 92%, greater than about 94%, greater than about 96%, greater than about 98%, or about 100%, or greater than about 65% and at most about 99%, or greater than about 70% and at most about 99%, or greater than about 75% and at most about 99%, or greater than about 80% and at most about 99%, or greater than about 85% and at most about 99%.

[0062] In cases where the percentage of cells expressing at least one additional marker in the enriched heterogeneous kidney cell population is determined, and the additional marker is or includes a method for determining podocyte protein, the percentage of cells expressing podocyte protein in the enriched heterogeneous kidney cell population that expresses therapeutic potential can be greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, greater than about 98%, or about 100%.

[0063] In cases where the percentage of cells expressing at least one additional marker in the enriched heterogeneous kidney cell population is determined, and the additional marker is or includes RACK1, the percentage of RACK1-expressing cells in the enriched heterogeneous kidney cell population can be greater than about 80%, greater than about 81%, greater than about 82%, greater than about 83%, greater than about 84%, greater than about 85%, greater than about 86%, greater than about 87%, greater than about 88%, greater than about 89%, greater than about 90%, greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, greater than about 99%, or about 100%.

[0064] In a method for determining the therapeutic potential of an enriched heterogeneous renal cell population, the determination may include not only determining the expression of at least one cell adhesion marker (at least two, at least three, or four of any one of ICAM5, MCAM, CNTNAP1, and / or MMP2) and optionally at least one renal-derived marker (at least two, at least three, at least four, or five of any one of SIX1, RET, OSR1, LHX1, and / or FGF8), but also optionally at least one additional cell adhesion marker (any one, or any two or three of COL11A1, PECAM1, and / or E-cadherin), and optionally one or more additional markers (e.g., TGFβ2, nephrotic protein, podocyte protein, and / or RACK-1), but optionally, it may further determine whether the cells of the enriched heterogeneous renal cell population secrete TGFβ2. In some cases, further measurement of TGFβ2 secretion from enriched heterogeneous kidney cell populations can identify these populations as having therapeutic potential. In some cases, measuring TGFβ2 secretion from enriched heterogeneous kidney cell populations can determine the amount of approximately TGFβ2 secreted by the cells. In such cases, further measurements can be taken for values ​​greater than approximately 1 ng TGFβ2 / million cells, greater than approximately 1.2 ng TGFβ2 / million cells, greater than approximately 1.4 ng TGFβ2 / million cells, greater than approximately 1.6 ng TGFβ2 / million cells, greater than approximately 1.8 ng TGFβ2 / million cells, greater than approximately 2.0 ng TGFβ2 / million cells, greater than approximately 2.2 ng TGFβ2 / million cells, greater than approximately 2.4 ng TGFβ2 / million cells, greater than approximately 2.6 ng TGFβ2 / million cells, greater than approximately 2.8 ng TGFβ2 / million cells, greater than approximately 3.0 ng TGFβ2 / million cells, greater than approximately 3.5 ng TGFβ2 / million cells, greater than approximately 4.0 ng TGFβ2 / million cells, greater than approximately 4.5 ng TGFβ2 / million cells, greater than approximately 5.0 ng TGFβ2 / million cells, and greater than approximately 5.5 ng TGFβ2 / million cells. TGFβ2 / million cells, greater than approximately 6.0 ng TGFβ2 / million cells, greater than approximately 6.5 ng TGFβ2 / million cells, greater than approximately 7.0 ng TGFβ2 / million cells, greater than approximately 7.5 ng TGFβ2 / million cells, greater than approximately 8.0 ng TGFβ2 / million cells, greater than approximately 8.5 ng TGFβ2 / million cells, greater than approximately 9.0 ng TGFβ2 / million cells, greater than approximately 9.5 ng TGFβ2 / million cells, greater than approximately 10.0 ng TGFβ2 / million cells, greater than approximately 11.0 ng TGFβ2 / million cells, greater than approximately 12.0 ng TGFβ2 / million cells, greater than about 13.0 ng TGFβ2 / million cells, greater than about 14.0 ng TGFβ2 / million cells, greater than about 15.0 ng TGFβ2 / million cells, or about 1 ng to about 40 ng TGFβ2 / million cells, about 1 ng to about 35 ng TGFβ2 / million cells, about 1 ng to about 30 ng TGFβ2 / million cells, about 1 ng to about 25 ng TGFβ2 / million cells, about 1 ng to about 20 ng TGFβ2 / million cells, about 2 ng to about 40 ng TGFβ2 / million cells, about 2 ng to about 35 ng TGFβ2 / million cells, about 2 ng to about 30 ng TGFβ2 / million cells, about 2 ng to about 25 ng TGFβ2 / million cells, about 2 ng to about 20 ng TGFβ2 / million cells, about 3 ng to about 40 ng TGFβ2 / million cells, approximately 3 ng to approximately 35 ng TGFβ2 / million cells, approximately 3 ng to approximately 30 ng TGFβ2 / million cells, approximately 3 ng to approximately 25 ng TGFβ2 / million cells, or approximately 3 ng to approximately 20 ng TGFβ2 / million cells, are secreted to identify enriched heterogeneous renal cell populations as having therapeutic potential. Cells can secrete the stated amounts of TGFβ2 / million cells over approximately 24 hours, approximately 30 hours, approximately 36 hours, approximately 42 hours, approximately 48 hours, approximately 54 hours, approximately 60 hours, approximately 66 hours, approximately 72 hours, approximately 78 hours, approximately 84 hours, approximately 90 hours, approximately 96 hours, approximately 102 hours, or approximately 108 hours, for example, in post-culture medium.

[0065] For example, in some cases of the method, if the TGFβ2 secretion of the enriched heterogeneous renal cell population is further measured, if the secretion per million cells is further measured at time intervals of approximately 18 hours, approximately 24 hours, approximately 30 hours, approximately 36 hours, approximately 20 hours to approximately 36 hours, approximately 18 hours to approximately 30 hours, approximately 18 to 24 hours, approximately 24 hours to approximately 36 hours, or approximately 24 hours to approximately 30 hours, greater than approximately 0.7 ng, greater than approximately 0.8 ng, greater than approximately 0.9 ng, greater than 1.0 ng, greater than approximately 1.1 ng, greater than approximately 1.2 ng, approximately 0.7 ng to approximately 1.75 ng, approximately 0.7 ng to approximately 1.5 ng, approximately 0.7 ng to approximately 1.25 ng, approximately 0.8 ng to approximately 1.75 ng, approximately 0.8 ng to approximately 1.5 ng, approximately 0.8 ng to approximately 1.25 ng, approximately 0.9 ng to approximately 1.75 ng, approximately 0.9 ng to approximately 1.5 ng, or approximately 0.9 ng to approximately 1.5 ng, within the time intervals of approximately 18 hours, approximately 24 hours, approximately 30 hours, approximately 36 hours, approximately 20 hours to approximately 36 hours, approximately 18 hours to approximately 30 hours, approximately 18 hours to approximately 24 hours, approximately 24 hours to approximately 36 hours, or approximately 24 hours to approximately 30 hours, greater than approximately 0.7 ng, greater than approximately 0.8 ng, greater than approximately 0.9 ng to approximately 1.5 ng, or approximately 0.9 ng to approximately 1.5 ng, greater than approximately 0.9 ng to approximately 1.5 ng, greater than approximately 0.9 TGFβ2 in amounts of approximately 0.9 ng to approximately 1.25 ng, approximately 1.0 ng to approximately 1.75 ng, or approximately 1.0 ng to approximately 1.5 ng can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0066] To give another example, if we further measure the cellular secretion of TGFβ2 from the enriched heterogeneous renal cell population, and if we further measure the secretion of greater than about 3.5 ng, greater than about 3.7 ng, greater than about 3.9 ng, greater than 4.1 ng, greater than about 4.3 ng, greater than about 4.5 ng, or about 3.5 ng to about 8.0 ng, about 3.5 ng to about 7.5 ng, about 3.5 ng to about 7.0 ng, about 3.9 ng to about 8.0 ng, about 3.9 ng to about 7.5 ng, or about 3.9 ng to about 7.5 ng, about 3.9 ng to about 7.0 ng, or about 3.9 ng to about 7.0 ng per million cells within time periods of approximately 36 hours, approximately 42 hours, approximately 48 hours, approximately 54 hours, approximately 60 hours, approximately 36 hours to about 60 hours, approximately 36 hours to about 54 hours, approximately 36 hours and about 48 hours, approximately 42 hours to about 60 hours, about 42 hours to about 54 hours, or about 42 hours to about 48 hours, within time periods of approximately 36 hours, approximately 42 hours to about 60 hours, approximately ... TGFβ2 in amounts of approximately 4.1 ng to approximately 8.0 ng, approximately 4.1 ng to approximately 7.5 ng, approximately 4.1 ng to approximately 7.0 ng, approximately 4.5 ng to approximately 8.0 ng, approximately 4.5 ng to approximately 7.5 ng, or approximately 4.5 ng to approximately 7.0 ng can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0067] In a further example, if the TGFβ2 secretion of the enriched heterogeneous renal cell population is further measured, and if the secretion per million cells is further measured at time intervals of approximately 60 hours, approximately 72 hours, approximately 78 hours, approximately 84 hours, approximately 60 hours to approximately 84 hours, approximately 60 hours to approximately 78 hours, approximately 60 hours to approximately 72 hours, approximately 72 hours to approximately 84 hours, or approximately 72 hours to approximately 78 hours, the secretion is greater than approximately 8.0 ng, greater than approximately 8.25 ng, greater than approximately 8.5 ng, greater than approximately 8.75 ng, greater than approximately 9.0 ng, approximately 8.0 ng to approximately 25 ng, approximately 8.0 ng to approximately 20 ng, approximately 8.0 ng to approximately 15 ng, approximately 8.25 ng to approximately 25 ng, approximately 8.25 ng to approximately 20 ng, approximately 8.25 ng to approximately 15 ng, approximately 8.5 ng to approximately 25 ng, or approximately 8.5 ng to approximately 20 ng, within the time intervals of approximately 60 hours, approximately 72 hours, approximately 78 hours, approximately 84 hours, approximately 60 hours to approximately 84 hours, approximately 60 hours to approximately 78 hours, approximately 60 hours to approximately 72 hours, approximately 72 hours to approximately 84 hours, or approximately 72 hours to approximately 78 hours, respectively, at a rate greater than approximately 8.0 ng, greater than approximately 8.25 ng, greater than approximately 8.5 ng to approximately 20 ng, or approximately 8.5 ng to approximately 20 ng, respectively, within the time intervals of approximately 60 hours, approximately 72 hours, approximately 78 hours, approximately 84 hours, approximately 60 hours to approximately 84 hours, approximately 60 hours to approximately TGFβ2 in doses of approximately 8.5 ng, approximately 15 ng, approximately 8.75 ng to approximately 25 ng, or approximately 8.75 ng to approximately 20 ng, or approximately 8.75 ng to approximately 15 ng, can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0068] In another method for identifying enriched heterogeneous kidney cell populations as having therapeutic potential, as provided in this disclosure, the enriched heterogeneous kidney cell population can be identified as having therapeutic potential by measuring TGFβ2 expression in the cells of the enriched heterogeneous kidney cell population. In these methods, the enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) more than about 50% of the cells in the enriched heterogeneous kidney cell population express TGFβ2, and / or (ii) TGFβ2 is secreted by the cells of the enriched heterogeneous kidney cell population at a rate of at least about 1.0 ng or about 2.0 ng / 1,000,000 cells in the enriched heterogeneous kidney cell population.

[0069] Among these other methods, the method of measuring TGFβ2 expression in enriched heterogeneous kidney cell populations can identify enriched heterogeneous kidney cell populations as having therapeutic potential if more than about 50% of the cells in the enriched heterogeneous kidney cell population are found to express TGFβ2. In some cases, if more than about 40% and up to about 85%, more than about 40% and up to about 80%, more than about 40% and up to about 75%, more than about 40% and up to about 70%, more than about 45% and up to about 85%, more than about 45% and up to about 80%, more than about 45% and up to about 75%, more than about 45% and up to about 70%, more than about 50% and up to about 85%, more than about 50% and up to about 80%, more than about 50% and up to about 75%, more than about 50% and up to about 70%, more than about 55% and up to about 70%, more than about 55% and up to about 85%, more than about 55% and up to about 80%, or more than about 55% and up to about 75% of the cells in the enriched heterogeneous kidney cell population are found to express TGFβ2, the enriched heterogeneous kidney cell population can be identified as having therapeutic potential. In some cases, if more than 40%, more than 42.5%, more than 45%, more than 47.5%, more than 50%, more than 52.5%, more than 55%, more than 57.5%, or more than 60% of the enriched heterogeneous kidney cell population is found to express TGFβ2, the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.

[0070] Among these other methods, the method of determining TGFβ2 expression in enriched heterogeneous kidney cell populations can identify enriched heterogeneous kidney cell populations as having therapeutic potential if the amount of TGFβ2 secreted by the enriched heterogeneous kidney cell populations is determined to be at least 1.0 ng / 1,000,000 enriched heterogeneous kidney cell populations. In some cases, if the enriched heterogeneous renal cell population secretes TGFβ2 in amounts greater than approximately 1 ng, or greater than approximately 1.2 ng, or greater than approximately 1.4 ng, or greater than approximately 1.6 ng, or greater than approximately 1.8 ng, or greater than approximately 2 ng, or greater than approximately 2.2 ng, or greater than approximately 2.4 ng, or greater than approximately 2.6 ng, or greater than approximately 2.8 ng, or greater than approximately 3.0 ng, or greater than approximately 3.5 ng, or greater than approximately 4.0 ng, or greater than approximately 4.5 ng, or greater than approximately 5.0 ng, or greater than approximately 5.5 ng, or greater than approximately 6.0 ng, or greater than approximately 6.5 ng, or greater than approximately 7.0 ng, or greater than approximately 7.5 ng, or greater than approximately 8.0 ng, or greater than approximately 8.5 ng, or greater than approximately 9.0 ng, or greater than approximately 9.5 ng, or greater than approximately 10 ng, or greater than approximately 11 ng, the amount of TGFβ2 secreted by the cells is considered significant. An enriched heterogeneous renal cell population of 10 ng, or more than about 12 ng, or more than about 13 ng, or more than about 14 ng, or more than about 15 ng / million cells can be identified as having therapeutic potential. Alternatively, if the enriched heterogeneous kidney cell population secretes TGFβ2 in amounts of about 1 ng to about 40 ng / million cells, or about 1 ng to about 35 ng / million cells, or about 1 ng to about 30 ng / million cells, or about 1 ng to about 25 ng / million cells, or about 1 ng to about 20 ng / million cells, or about 2 ng to about 40 ng / million cells, or about 2 ng to about 35 ng / million cells, or about 2 ng to about 30 ng / million cells, or about 2 ng to about 25 ng / million cells, or about 2 ng to about 20 ng / million cells, or about 3 ng to about 40 ng / million cells, or about 3 ng to about 35 ng / million cells, or about 3 ng to about 30 ng / million cells, or about 3 ng to about 25 ng / million cells, or about 3 ng to about 20 ng / million cells, then the enriched heterogeneous kidney cell population can be identified as having therapeutic potential.The TGFβ2 secreted by the enriched heterogeneous kidney cell population in these amounts can, for example, in the culture medium, within a time period of about 24 hours, about 30 hours, about 36 hours, about 42 hours, about 48 hours, about 54 hours, about 60 hours, about 66 hours, about 72 hours, about 78 hours, about 84 hours, about 90 hours, about 96 hours, about 102 hours, about 108 hours, or about 24 hours to about 108 hours, or about 36 hours to about 108 hours, or about 24 hours to about 90 hours, or about 24 hours to about 72 hours, or about 48 hours to about 72 hours.

[0071] In the case of these other methods, the method of measuring TGFβ2 expression in enriched heterogeneous kidney cell populations is used if the enriched heterogeneous kidney cell populations secrete greater than about 0.7 ng, greater than about 0.8 ng, greater than about 0.9 ng, greater than 1.0 ng, greater than about 1.1 ng, greater than about 1.2 ng, greater than 1.4 ng, greater than 1.5 ng, greater than 2.0 ng, greater than 2.5 ng per million cells within time periods of about 18 hours, about 24 hours, about 30 hours, about 36 hours, about 20 hours to about 36 hours, about 18 hours to about 30 hours, about 18 hours to about 24 hours, about 24 hours to about 36 hours, or about 24 hours to about 30 hours. Approximately 0.9 ng to 1.75 ng, approximately 0.9 ng to 1.5 ng, approximately 0.9 ng to 1.25 ng, approximately 1.0 ng to 1.75 ng, approximately 1.0 ng to 1.5 ng, approximately 4.0 ng, approximately 1.0 ng to 5.0 ng, approximately 1.0 ng to 6.0 ng, approximately 1.0 ng to 7.0 ng, approximately 2.0 ng to 4.0 ng, approximately 2.0 ng to 5.0 ng, approximately 2.0 ng to 6.0 ng, or approximately 2.0 ng to 7.0 ng of TGFβ2 can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0072] In other cases of these methods, if the enriched heterogeneous renal cell population is measured to secrete greater than about 3.5 ng, greater than about 3.7 ng, greater than about 3.9 ng, greater than 4.1 ng, greater than about 4.3 ng, greater than about 4.5 ng, about 3.5 ng to about 8.0 ng, about 3.5 ng to about 7.5 ng, about 3.5 ng to about 7.0 ng, about 3.9 ng to about 8.0 ng, about 3.9 ng to about 7.5 ng, about 3.9 ng to about 7.0 ng, about 4.1 ng to about 8.0 ng, or about 4.1 ng to about 8.0 ng per million cells within time periods of about 36 hours, about 42 hours, about 48 hours, about 54 hours, about 60 hours, about 36 hours to about 60 hours, about 36 hours to about 54 hours, about 36 hours to about 48 hours, about 42 hours to about 60 hours, about 42 hours to about 54 hours, or about 42 hours to about 48 hours, the following parameters are also considered: Approximately 4.1 ng to approximately 7.5 ng, approximately 4.1 ng to approximately 7.0 ng, approximately 4.5 ng to approximately 8.0 ng, approximately 4.5 ng to approximately 7.5 ng, or approximately 4.5 ng to approximately 7.0 ng TGFβ2 can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0073] In another instance of these methods, if the enriched heterogeneous renal cell population is measured to secrete greater than approximately 8.0 ng, greater than approximately 8.25 ng, greater than approximately 8.5 ng, greater than approximately 8.75 ng, greater than approximately 9.0 ng, approximately 8.0 ng to approximately 25 ng, approximately 8.0 ng to approximately 20 ng, approximately 8.0 ng to approximately 15 ng, approximately 8.25 ng to approximately 25 ng, approximately 8.25 ng to approximately 20 ng, approximately 8.25 ng to approximately 15 ng, approximately 8.5 ng to approximately 25 ng, approximately 8.5 ng to approximately 20 ng, or approximately 8.5 ng to approximately 20 ng, per million cells within time periods of approximately 60 hours, approximately 72 hours, approximately 78 hours, approximately 84 hours, approximately 60 hours to approximately 84 hours, approximately 60 hours to approximately 78 hours, approximately 60 hours to approximately 72 hours, approximately 72 hours to approximately 84 hours, or approximately 72 hours to approximately 78 hours, the percentage of cells secreting is also considered. Approximately 15 ng, 8.75 ng to 25 ng, 8.75 ng to 20 ng, or 8.75 to 15 ng of TGFβ2 can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0074] Among these other methods, the method of determining TGFβ2 expression in enriched heterogeneous kidney cell populations can identify them as having therapeutic potential if both of the following conditions are met: (i) approximately a certain percentage of cells in the population express TGFβ2 and (ii) the cells in the enriched heterogeneous kidney cell population secrete approximately a certain amount of TGFβ2. In some cases, enriched heterogeneous kidney cell populations can be identified as having therapeutic potential if (i) more than approximately 50% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) TGFβ2 is secreted at a level of at least approximately 1.0 ng, 1.5 ng, 2.0 ng, 2.5 ng, 3.0 ng, 3.5 ng, 4.0 ng, 4.5 ng, or 5.0 ng / million cells. In some cases, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) approximately 50% to approximately 80% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) TGFβ2 is secreted at a level of at least approximately 1.0 ng, 1.5 ng, 2.0 ng, 2.5 ng, 3.0 ng, 3.5 ng, 4.0 ng, 4.5 ng, or 5.0 ng / million cells. In some cases, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) more than approximately 55% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) TGFβ2 is secreted at a level of at least approximately 1.0 ng, 1.5 ng, 2.0 ng, 2.5 ng, 3.0 ng, 3.5 ng, 4.0 ng, 4.5 ng, or 5.0 ng / million cells. In some cases, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) approximately 55% to approximately 75% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) TGFβ2 is secreted at a level of at least approximately 1.0 ng, 1.5 ng, 2.0 ng, 2.5 ng, 3.0 ng, 3.5 ng, 4.0 ng, 4.5 ng, or 5.0 ng / million cells. In some cases, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) more than approximately 50% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) TGFβ2 is secreted at a level of at least approximately 1.0 ng to approximately 50 ng, or approximately 2.0 ng to approximately 40 ng, or approximately 3.0 ng to approximately 30 ng / million cells. In some cases, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) approximately 50% to approximately 80% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) TGFβ2 is secreted at a rate of approximately 1.0 ng to approximately 50 ng, or approximately 2.0 ng to approximately 40 ng, or approximately 3.0 ng to approximately 30 ng / million cells.In some cases, an enriched heterogeneous kidney cell population can be identified as having therapeutic potential if (i) more than about 55% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) TGFβ2 is secreted at a rate of about 1.0 ng to about 50 ng, or about 2.0 ng to about 40 ng, or about 3.0 ng to about 30 ng / million cells. In these cases, the amount of TGFβ2 secreted by cells can be secreted over a period of approximately 24 hours to approximately 120 hours, or approximately 24 hours, approximately 36 hours, approximately 24 hours to approximately 72 hours, approximately 48 hours to approximately 120 hours, approximately 48 hours to approximately 96 hours, approximately 48 hours, approximately 72 hours, approximately 96 hours, or approximately 120 hours.

[0075] For example, among these other methods, a method for determining TGFβ2 expression in enriched heterogeneous kidney cell populations can identify a population with therapeutic potential if it measures that: (i) more than about 50%, or more than about 55%, or about 50% of the cells in the enriched heterogeneous kidney cell population express TGFβ2 and (ii) secrete TGFβ2, wherein TGFβ2 is secreted in amounts of at least about the following: (a) 0.7 ng / 1,000,000 enriched heterogeneous kidney cell populations over a period of about 24 hours; (b) 3.5 ng / 1,000,000 enriched heterogeneous kidney cell populations over a period of about 48 hours; and / or (c) 8.0 ng / 1,000,000 enriched heterogeneous kidney cell populations over a period of about 72 hours. In some of these instances, the amount of TGFβ2 secreted over a period of approximately 24 hours may be at least about 1.0 ng, at least about 1.5 ng, at least about 2.0 ng, or at least about 2.5 ng / 1,000,000 enriched heterogeneous kidney cell populations.

[0076] Among these other methods, the method of determining TGFβ2 expression in enriched heterogeneous kidney cell populations can also determine the expression of at least one kidney-derived marker, wherein the at least one kidney-derived marker is one or more of SIX2, RET, OSR1, LHX1, or FGF8. Determining further expression of at least one kidney-derived marker (in addition to TGFβ2) in enriched heterogeneous kidney cell populations can identify enriched heterogeneous kidney cell populations as having therapeutic potential.

[0077] The at least one renal biomarker that can be further measured in the method may be any one of SIX1, RET, OSR1, LHX1, or FGF8. The at least one renal biomarker that can be further measured in the method may be or may include any two of the following renal biomarkers: (a) SIX2 and OSR1; (b) SIX2 and LHX1; (c) SIX2 and RET; (d) SIX2 and FGF8; (e) OSR1 and LHX1; (f) OSR1 and RET; (g) OSR1 and FGF8; (h) LHX1 and RET; (i) LHX1 and FGF8; (j) RET and FGF8. The at least one renal biomarker that can be further determined in the method may be or may include any one of the following three renal biomarkers: (a) SIX2, OSR1 and LHX1, (b) SIX2, OSR1 and RET, (c) SIX2, OSR1 and FGF8, (d) SIX2, LHX1 and RET, (e) SIX2, LHX1 and FGF8, (f) SIX2, RET and FGF8, (g) OSR1, LHX1 and RET, (h) OSR1, LHX1 and FGF8, (i) OSR1, RET and FGF8, or (j) LHX1, RET and FGF8. The at least one renal biomarker that can be further measured in the method may be or may include any one of the following four renal biomarkers: (a) SIX2, OSR1, LHX1, and RET; (b) SIX2, OSR1, LHX1, and FGF8; (c) SIX2, LHX1, RET, and FGF8; (d) SIX2, OSR1, RET, and FGF8; or (e) OSR1, LHX1, RET, and FGF8. The at least one renal biomarker that can be further measured in the method may include all five renal biomarkers: SIX2, OSR1, LHX1, RET, and FGF8.

[0078] In these methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, the expression of at least one renal biomarker (in addition to TGFβ2 expression) is further determined. The determination of at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers may further include determining the percentage of cells in the enriched heterogeneous renal cell population expressing at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers. If the percentage of cells in the enriched heterogeneous renal cell population expressing at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers is further determined, then if approximately a certain or specific percentage of cells in the enriched heterogeneous renal cell population express at least one (or at least two, or at least three, or at least four, or all five) renal biomarkers, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. A certain, specific percentage of cells in a heterogeneous population of kidney cells enriched by any one of at least one (at least one, or at least two, or at least three, or at least four, or five) kidney-derived markers can be any percentage of any one of the kidney-derived markers discussed earlier herein.

[0079] In these other methods, the expression of TGFβ2 in the enriched heterogeneous renal cell population is measured, and optionally the expression of at least one renal biomarker is measured. The expression of at least one additional biomarker can be further measured to identify the enriched heterogeneous renal cell population as having therapeutic potential. Where at least one additional biomarker can be further measured, this additional biomarker can be at least one (at least two, at least three, or four) selected from gamma-glutamyltranspeptidase (GGT)-1, cytokeratin (CK)18, vascular endothelial growth factor (VEGF), or kidney injury molecule (KIM)-1. At least one added marker may be GGT-1, or may be CK18, or may be VEGF, or may be KIM-1, or may be GGT-1 and CK18, or may be GGT-1 and VEGF, or may be GGT-1 and KIM-1, or may be CK-18 and VEGF, or may be CK18 and KIM-1, or may be VEGF and KIM-1, or may be GGT-1, CK18 and VEGF, or may be GGT-1, VEGF and KIM-1, or may be CK-18, VEGF and KIM-1, or may be GGT-1, CK-18 and KIM-1, or may be GGT-1, CK18, VEGF and KIM-1.

[0080] Further expression of at least one additional biomarker in enriched heterogeneous kidney cell populations can identify them as having therapeutic potential.

[0081] In some cases, determining whether the enriched heterogeneous kidney cell population further expresses at least one additional marker may involve determining whether a certain percentage of the enriched heterogeneous kidney cell population expresses at least one additional marker, or determining whether the enriched heterogeneous kidney cell population secretes at least one additional marker. Determining whether a certain percentage of the enriched heterogeneous kidney cell population expresses at least one additional marker and / or whether the enriched heterogeneous kidney cell population secretes at least one additional marker can identify the enriched heterogeneous kidney cell population as having therapeutic potential.

[0082] In cases where at least one added biomarker is GGT-1, a further determination of the percentage of cells expressing GGT-1 in the enriched heterogeneous kidney cell population can be made to identify the enriched heterogeneous kidney cell population as having therapeutic potential. In these cases, the percentage of GGT-1-expressing cells in the enriched heterogeneous kidney cell population that can be identified as having therapeutic potential can be greater than about 4%, greater than about 10%, greater than about 15%, greater than about 20%, greater than about 25%, greater than about 30%, greater than about 35%, greater than about 40%, greater than about 45%, or greater than about 50%. Alternatively, the percentage of GGT-1-expressing cells in the enriched heterogeneous kidney cell population that can be identified as having therapeutic potential can be about 4% to 85%, about 4% to 82%, about 4.5% to 80%, about 4.5% to 75%, or about 4.5% to 70%.

[0083] In cases where at least one added biomarker is CK18, a further determination of the percentage of cells expressing CK18 in the enriched heterogeneous kidney cell population can be made to identify the enriched heterogeneous kidney cell population as having therapeutic potential. In these cases, the percentage of CK18-expressing cells that can identify the enriched heterogeneous kidney cell population as having therapeutic potential can be greater than about 80%, greater than about 82%, greater than about 84%, greater than about 86%, greater than about 88%, greater than about 90%, greater than about 92%, or greater than about 94%.

[0084] In the case of at least one added biomarker being VEGF and / or KIM-1, the cellular secretion of VEGF and / or KIM-1 by the enriched heterogeneous kidney cell population can be further measured to identify the enriched heterogeneous kidney cell population as having therapeutic potential.

[0085] It should be understood that if the percentage of cells expressing a certain biomarker is provided as "about" a specific number, such as about 5%, the percentage of cells does not need to be exactly that specific number, for example, exactly 5%. More precisely, it should be understood that if the percentage of cells expressing a certain biomarker is provided as "about" a specific number, such as about 5%, then the percentage of cells expressing the certain biomarker may be within 10% of that specific number, for example, between 4.5% and 5.5%. It should also be understood that if cells express a certain biomarker by secreting an amount of "about" a specific number, such as about 5 ng, that amount does not need to be exactly that specific number, for example, exactly 5 ng. More precisely, it should be understood that the amount of biomarker secreted may be within 10% of that specific number, for example, between 4.5% and 5.5 ng.

[0086] In any of the methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, determining whether the cells in the enriched heterogeneous renal cell population express any marker, such as cell adhesion markers (ICAM5, MCAM, CNTNAP1, MMP2), renal markers (SIX1, RET, OSR1, LHX1, or FGF8), other cell adhesion markers (COL11A1, PECAM1, or E-cadherin), additional markers (e.g., TGFβ2, nephrotic protein, podocyte protein, or RACK-1), TGFβ2, or added markers (GGT1, CK18, VEGF, or KIM-1), can be done by detecting markers in the form of nucleic acids (e.g., mRNA or miRNA) or peptides, and can be done by any suitable assay / technique. For example, if expression is determined in peptide form, it can be determined by assays such as Western blot, fluorescence-activated cell sorting (FACS), or enzyme-linked immunosorbent assay (ELISA). If expression is measured in nucleic acid form, it can be determined by assays such as Southern blotting, polymerase chain reaction (PCR) or reverse transcriptase PCR, gene expression serial analysis (SAGE), Mass ARRAY, or fluorescence in situ hybridization (FISH). Expression can be determined by batch or single-cell assays, or after collecting conditioned medium from cell samples enriched with heterogeneous kidney cell populations. The assay can be a assay using labeled detection reagents. The labeled detection reagents can include (a) a portion directly or indirectly combined with the marker and (b) a detection portion. Non-restrictive detection portions include radioactive isotopes, for example, 35 S, 14 C, 125 I, 3 H and 131I. Colloidal gold particles, fluorescent labels such as Texas Red, Rhodamine, fluorescein, dansyl, Lissamine, phycoerythrin, phycocyanin, SPECTRUM ORANGE, SPECTRUM GREEN1, and enzyme substrates such as firefly luciferase, bacterial luciferase, luciferin, horseradish peroxidase, alkaline phosphatase, or β-galactosidase.

[0087] The enriched heterogeneous renal cell population identified as having therapeutic potential in any of the methods may be enriched with one or more renal cell types, such as renal epithelial cells, renal tubular cells, renal tubular epithelial cells, or proximal renal tubular cells. Enrichment of the heterogeneous renal cell population for these one or more renal cell types can refer to the enriched heterogeneous renal cell population having a greater percentage of one or more renal cell types than the patient's renal tissue, the patient's renal biopsy, or an in vitro culture of cells established from the patient's renal tissue or renal biopsy (which may be collectively referred to as the "initial renal cell population"). If the in vitro culture of cells established from the patient's renal tissue or the patient's renal biopsy is used, the initial renal cell population may be a renal cell preparation containing dissociated cells from the renal tissue or renal biopsy (e.g., cells dissociated from the renal tissue or renal biopsy via chopping and / or enzymatic digestion), which may have been treated or may not have been treated to remove red blood cells and debris. In addition to being rich in renal epithelial cells, renal tubular cells, renal tubular epithelial cells and / or proximal tubular cells, the enriched heterogeneous renal cell population may also include other renal cell types, such as glomerular cells, podocytes, collecting duct cells and / or vascular cells.

[0088] Because the enriched heterogeneous kidney cell population has been prepared from the initial kidney cell population (e.g., patient's kidney tissue, patient's renal biopsy, or an in vitro culture of cells established from patient's kidney tissue or renal biopsy) via a method including a separation step, the enriched heterogeneous kidney cell population may be rich in one or more kidney cell types. The separation step may be a step based on the buoyancy density of the cells, separating cells from the initial kidney cell population that has been passaged no more than once, twice, or three times. If the separation step is based on the buoyancy density of the cells, the separation step may utilize a single-step or multi-step continuous or discontinuous density gradient using a density gradient medium such as glycerol, glucose OptiPrep, Percoll, or Ficoll-Paque. Using such a density gradient medium in this manner can result in the separation of cells from the initial kidney cell population (or an initial kidney cell population that has been passaged no more than once, twice, or three times) into one or more distinguishable fractions from which the enriched heterogeneous kidney cell population can be clearly identified and separated. A distinguishable fraction can be a fraction / those fractions in which the buoyant density of cells is greater than about 1.045 g / mL, or greater than 1.045 g / mL, or greater than or equal to 1.045 g / mL. A distinguishable fraction can be a fraction / those fractions in which the buoyant density of cells is greater than about 1.04 g / mL, or greater than 1.04 g / mL, or greater than or equal to 1.04 g / mL, or greater than about 1.0419 g / mL, or greater than 1.0419 g / mL, or greater than or equal to 1.0419 g / mL. A distinguishable fraction can be a fraction / those fractions in which the buoyant density is from about 1.045 g / mL to about 1.091 g / mL, or from about 1.045 g / mL to about 1.052 g / mL. Alternatively, the separation step can be based on whether cells from the initial kidney cell population (or cells from an initial kidney cell population that has been passaged no more than once, twice, or three times) express a specific marker on their surface. If the separation step is based on the expression of a specific cell surface marker, then the separation step can be a separation step using flow cytometry. If the cells express a specific surface marker characteristic of, for example, renal epithelial cells, renal tubular cells, renal tubular epithelial cells, or proximal tubular cells, such as nephrotic protein or cytokeratin (CK), for example, CK18, then flow cytometry can sort cells from the initial kidney cell population (or an initial kidney cell population that has been passaged no more than once, twice, or three times) to form a separated, enriched heterogeneous kidney cell population.

[0089] Prior to the separation step, enriched heterogeneous kidney cell populations prepared from the starting kidney cell population (or starting kidney cell populations that have been passaged one, two, or three times) can be cultured under hypoxic conditions. If cells are cultured under hypoxic conditions prior to the separation step, they can be cultured under conditions in which oxygen levels are less than about 20%, or less than about 15%, or less than about 10%, or less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%. If cells are cultured under hypoxic conditions, the cells can be cultured under hypoxic conditions for at least 6 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 14 hours, at least 16 hours, at least 20 hours, at least 24 hours, at least 30 hours, at least 36 hours, at least 42 hours, at least 48 hours, about 6 hours to about 48 hours, about 6 hours to about 36 hours, about 6 hours to about 24 hours, about 12 hours to about 48 hours, about 12 hours to about 36 hours, or about 12 hours to about 24 hours.

[0090] Typically, enriched heterogeneous kidney cell populations can be prepared from any starting cell population, such as in vitro cultures of cells established from a patient's kidney tissue or renal biopsy. If the enriched heterogeneous kidney cell population is prepared from an in vitro culture of cells established from a patient's kidney tissue or renal biopsy, the cells in the in vitro culture can be expanded by passage at most one, two, or three times. Alternatively, if desired, cells from an in vitro culture of cells established from kidney tissue or renal biopsy can be cryopreserved and then expanded by passage at most one, two, or three times. Once the cells have been expanded, the expanded cells can be cryopreserved. The expanded cells, whether cryopreserved or not, can then undergo a separation step, or can then be cultured under hypoxic conditions followed by a separation step. The enriched heterogeneous kidney cell population is separable by performing a separation step. Once the enriched heterogeneous kidney cell population has been separated, it can be frozen and / or analyzed before being used as a therapeutic agent.

[0091] If the enriched heterogeneous renal cell population is identified as having therapeutic potential according to any of the methods disclosed herein, it may be included in a pharmaceutical composition, administered in a method of treating a patient with renal disease in need of such treatment, and / or used to prepare a medicament for treating renal disease. If the enriched heterogeneous renal cell population is identified as having therapeutic potential and is included in a pharmaceutical composition, it may be formulated into a hydrogel composition or a liquid composition. The pharmaceutical composition may or may not include hyaluronic acid.

[0092] If the pharmaceutical composition is formulated into a hydrogel composition, the cells of the enriched heterogeneous kidney cell composition can be combined with a temperature-sensitive cell-stabilizing biomaterial. The temperature-sensitive cell-stabilizing biomaterial can be a biomaterial that is in a gel state at certain temperatures and in a liquid state at other temperatures. For example, if the biomaterial is temperature-sensitive, it may be in a gel state at or below about 8°C, substantially liquid at or above ambient temperature, and in a solid-liquid transition state between about 8°C and about ambient temperature; or in a gel state at or below about 4°C, liquid at or above about 37°C, and in a solid-liquid transition state between about 8°C and about 18°C; or in a gel state at or below about 2°C, liquid at or above about 37°C, and in a solid-liquid transition state between about 8°C and about 18°C; or in a gel state at or below about 2°C, and liquid at or above about 37°C; or in a gel state at or below about 4°C, and liquid at or above about 34°C; or in a gel state at or below about 6°C, and liquid at or above about 32°C. Temperature-sensitive cell-stabilizing biomaterials may include, or be made from, one or more naturally derived or recombinant proteins or peptides. Naturally derived or recombinant proteins or peptides may be recombinant extracellular matrix proteins, or extracellular matrix derived from the kidney or another tissue or organ, or gelatin. If the temperature-sensitive cell-stabilizing biomaterial is or includes gelatin, the gelatin may be derived from type I, collagen αI, such as porcine type I, collagen αI, or recombinant human type I, collagen αI. If the temperature-sensitive cell-stabilizing biomaterial is or includes gelatin, the gelatin may be present in the therapeutic composition at about 0.5% to about 1% by weight / volume (w / v), or about 0.8% to about 0.9% (w / v), or about 0.75% (w / v), or about 0.88% (w / v). The enriched heterogeneous kidney cell population may be dispersed throughout the biomaterial or substantially uniformly distributed throughout the biomaterial. The enriched heterogeneous kidney cell population may be formulated in the biomaterial, such as gelatin, such that the number of cells per mL of biomaterial is about 20 x 10⁻⁶. 6 cells / mL, approximately 40 x 10 6 cells / mL, approximately 60 x 10 6 cells / mL, approximately 100 x 10⁻⁶ 6 cells / mL, approximately 120 x 10 6 cells / mL, approximately 140 x 10 6 cells / mL, approximately 160 x 10 6 cells / mL, approximately 180 x 10 6 1 cell / mL or approximately 200 x 10 6Cells / mL.

[0093] If the pharmaceutical composition is formulated as a liquid composition, the enriched heterogeneous kidney cell population can be combined with any suitable liquid, such as a suitable cell storage or culture medium, saline, or a combination thereof, for immediate use or for cryopreservation until its use. If the therapeutic composition is a liquid composition, the cells of the enriched heterogeneous kidney cell population can be suspended in a pharmaceutically acceptable carrier or excipient, such as saline, buffered saline, dextran, water, polyethylene glycol, and / or any combination thereof. The cells of the enriched heterogeneous kidney cell population can be combined with a suitable liquid, such as a cell storage or culture medium, such that the number of cells per mL of liquid is approximately 20 x 10⁻⁶. 6 cells / mL, approximately 40 x 10 6 cells / mL, approximately 60 x 10 6 cells / mL, approximately 100 x 10⁻⁶ 6 cells / mL, approximately 120 x 10 6 cells / mL, approximately 140 x 10 6 cells / mL, approximately 160 x 10 6 cells / mL, approximately 180 x 10 6 1 cell / mL or approximately 200 x 10 6 Cells / mL.

[0094] If an enriched heterogeneous renal cell population is identified as having therapeutic potential, the enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population may be administered to a patient in a method of treating kidney disease or may be used in a method of treating kidney disease. If an enriched heterogeneous renal cell population is identified as having therapeutic potential, the enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population may be used to prepare a medicament for treating kidney disease. Kidney disease can be acute or chronic renal failure of any stage or degree. Kidney disease can originate in the kidneys or it can be secondary to another condition, such as heart failure, hypertension, diabetes, autoimmune disease, or liver disease. Alternatively, kidney disease can be caused by acute damage to the kidneys or it can be a result of renal and / or urinary tract abnormalities. Kidney disease may further include endocrine dysfunctions such as anemia, such as erythropoietin deficiency, and mineral imbalances, such as vitamin D deficiency.

[0095] If an enriched heterogeneous renal cell population is identified as having therapeutic potential, treatment of kidney disease can be achieved by administering the enriched heterogeneous renal cell population or a pharmaceutical composition containing such an enriched heterogeneous renal cell population. Kidney disease can be treated by restoring, stabilizing, improving, reducing renal fibrosis, or reducing renal inflammation in the kidneys of patients requiring such treatment. Treatment of kidney disease can also involve restoring mineral balance, electrolyte balance, fluid homeostasis, reabsorption of essential nutrients, or metabolism of cysteine ​​protease inhibitor C in patients requiring such treatment, or alleviating anemia. Treatment of kidney disease can delay or prevent the need for dialysis in patients requiring treatment, or it can delay or prevent the need for a kidney transplant. If treatment of kidney disease delays a patient's need for dialysis or a kidney transplant, the delay can be at least 1 year, at least 1.5 years, at least 2 years, at least 2.5 years, at least 3 years, at least 3.5 years, at least 4 years, at least 4.5 years, at least 5 years, at least 5.5 years, at least 6 years, at least 6.5 years, at least 7 years, at least 7.5 years, at least 8 years, at least 8.5 years, at least 9 years, at least 9.5 years, or at least 10 years. Treatment of kidney disease can be assessed by observing improvements in a patient's serum albumin, albumin / globulin ratio (A / G ratio), serum phosphorus, serum sodium, kidney size (which can be measured by ultrasound), serum calcium, phosphorus:calcium ratio, serum potassium, proteinuria, urinary creatinine, serum creatinine, blood nitrogen urea (BUN), cholesterol levels, triglyceride levels, glomerular filtration rate (GFR), weight, blood pressure (mean general blood pressure, diastolic blood pressure, or systolic blood pressure), and physical endurance.

[0096] If an enriched heterogeneous renal cell population is identified as having therapeutic potential, it can be administered to a patient via any suitable route of administration known in the art. For example, an enriched heterogeneous renal cell population or a pharmaceutical composition comprising such an enriched heterogeneous renal cell population can be administered systemically to a patient requiring treatment for kidney disease. The enriched heterogeneous renal cell population or a pharmaceutical composition comprising such an enriched heterogeneous renal cell population can be administered at or within the kidney of a patient requiring treatment for kidney disease. If the enriched heterogeneous renal cell population is administered at or within the kidney of a patient requiring treatment for kidney disease, it can be administered via a single or multiple injections. It can be administered via direct laparotomy, via direct laparoscopy, transabdominally, or percutaneously. The enriched heterogeneous renal cell population or a pharmaceutical composition comprising such an enriched heterogeneous renal cell population can be administered by percutaneous injection into the renal cortex, or by percutaneous insertion of a guiding cannula to puncture the renal capsule and then injecting the enriched heterogeneous renal cell population into the kidney. An enriched heterogeneous renal cell population or a pharmaceutical composition containing such an enriched heterogeneous renal cell population can be administered by injection into the renal cortex of one or both kidneys of a patient. Administration of the enriched heterogeneous renal cell population or the pharmaceutical composition containing such an enriched heterogeneous renal cell population can be performed via two injections, wherein the first injection is administered into the renal cortex of one kidney of the patient, and the second injection is administered into the renal cortex of the other kidney of the patient.

[0097] Enriched heterogeneous renal cell populations or pharmaceutical compositions containing such enriched heterogeneous renal cell populations may be administered at a therapeutically effective dose via any suitable route. A therapeutically effective dose or amount for administration to a patient requiring treatment for kidney disease may include approximately 1-9 x 10⁻⁶ cells / day. 6 The enriched heterogeneous renal cell population is measured per gram of estimated patient kidney weight. A therapeutically effective dose of the drug composition may be approximately 1.0 x 10⁻⁶ cells / gram. 6 Approximately 2.0 x 10 6 Approximately 3.0 x 10 6 Approximately 4.0 x 10 6 Approximately 5.0 x 10 6 Approximately 6.0 x 10 6 Approximately 7.0 x 10 6 Approximately 8.0 x 10 6 Approximately 9.0 x 10 6 Approximately 2.0–7.0 x 10 6 Approximately 4.0–7.0 x 10 6 Or approximately 5.0 x 10 6 –7.0 x 10 6 The estimated patient kidney weight is calculated as the number of cells per gram of enriched heterogeneous kidney cell populations.

[0098] In methods of treating kidney disease, administration of a therapeutic composition comprising heterogeneous renal cell populations to a patient can be performed via first and second injections. The first and second injections can be administered approximately 3 to 12 months apart. Alternatively, the first and second injections can be administered approximately 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months apart. Furthermore, the first and second injections can be administered approximately 3 to 6 months apart, approximately 6 to 9 months apart, approximately 9 to 12 months apart, or approximately 3 to 9 months apart, approximately 6 to 9 months apart, or approximately 6 to 12 months apart.

[0099] Those skilled in the art will recognize or be able to determine many equivalents of the specific embodiments described herein using only conventional experiments. Such equivalents are intended to be covered by the appended claims.

[0100] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference to the same extent that each individual publication, patent or patent application is specifically and individually indicated to be incorporated herein by reference in its entirety. Example

[0101] Example 1 - Biomarkers expressed by SRC population cells elucidate their role in stabilizing and restoring kidney function in a CKD model. The pathways that drive tubulogenic activity in the function

[0102] introduce. Selected renal cells (SRCs) are examples of enriched heterogeneous renal cell populations, such as those rich in renal epithelial cells. SRCs are being advanced as an autologous cell-based therapy for the treatment of chronic kidney disease (CKD). The bioactivity of SRCs is partly attributed to their perceived ability to influence the signaling cascades mediating nephroptosis or regeneration within the kidney; and to express certain markers expressed by renal cells (e.g., ureteral buds and cap mesenchymal cells) during kidney development or nephroptosis. Studies are being conducted to determine whether human SRCs express other markers, such as cell adhesion markers, or secrete other factors, which further elucidates the mechanisms by which they stabilize and restore renal function in CKD models.

[0103] Methods / Detection of biomarker expression in SRC Human SRC (National Disease Research Interchange) (i) was used to perform mRNA-seq to identify differentially expressed genes (DEG; DEG relative to the source biopsy; P) adj(ii) scRNA-seq was performed to map gene expression, and (iii) miRNA-seq was performed for analysis. SRC markers were identified using Gene Ontology (GO) and / or the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and their networks were visualized using miRNet.ca.

[0104] Methods / scRNA-seq for mapping gene expressionSRC aliquots were stained with acridine orange and propidium iodide, and cell viability, concentration, and singleness were assessed using a LUNA-FX7 dual-fluorescence cell counter (Logos Biosystems). Cells were then treated using a 10x Genomics Chromium Controller and a Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3.1 Dual Index Kit (PN-1000268) according to the manufacturer's user guide (https: / / tinyurl.com / 4855859x). In short, approximately 5,000 cells / sample were loaded onto a Chromium chip G, with a target recovery of 3,000 cells / sample for library preparation. Single cells, reverse transcription reagents, and gel beads coated with barcoded oligomers were encapsulated together in oil droplets to generate gel beads (GEMs) in an emulsion. Reverse transcription was performed using a C1000 thermal cycler (Bio-Rad) to generate a complementary DNA (cDNA) library tagged with cell barcodes and a unique molecular index (UMI). GEM images were then disrupted, and the cDNA was purified using Dynabeads MyOneSILANE beads (Invitrogen) followed by 12 cDNA amplification cycles. The amplified cDNA library was purified using SPRIselect magnetic beads (Beckman Coulter) and quantified using an Agilent Bioanalyzer HighSensitivity DNA array (Agilent Technologies). Fragmentation, end repair, A-tailing, and double-sided size selection using SPRIselect beads were then performed. Illumina-compatible adaptors were ligated to the size-selected cDNA fragments. The adaptor-ligated cDNA fragments were then purified using SPRIselect beads. A uniquely identifiable index was added to the cDNA during the 12 amplification cycles. The completed sequencing libraries were then purified using SPRIselect beads, visualized using a Bioanalyzer High Sensitivity DNA microarray, and merged at an equimolar ratio. The merged libraries were then sequenced using an Illumina NextSeq 2000 system at the UNC High Throughput Sequencing Facility.The library was denatured and diluted according to the standard Illumina protocol, incorporating 1% PhiX sequencing control (Illumina), and sequenced in a P3 flow cell with paired ends (read 1: 28 cycles, i7 index: 10 cycles, i5 index: 10 cycles, read 2: 90 cycles) to a total depth of 1.2 billion read pairs through a quality filter. Demultiplexing and preliminary analysis were performed using Cell Ranger 7.1.0 with default settings. Analysis was performed using 10X Genomics' LoupeBrowser (Tables 1 and 2) or Seurat (Seurat, v5.0.1; Table 5).

[0105] Methods / Flow cytometry for detecting protein expression and cell surface staining Wash the SRC with flow buffer and resuspend it in flow buffer to 10 μL. 6 The concentration is [value missing]. Transfer 100 μl of cells resuspended in flow buffer to the wells of a 96-well round-bottom plate or 5 mL of 12x75 mm plastic tubes. If necessary, add an appropriate amount of Fc block to the wells / tubes for incubation at 4°C for 15 minutes. Next, add the conjugated monoclonal antibodies (clone WM59 for PECAM-1 (also known as CD31) detection, clone HCD54 for ICAM-1 (also known as CD54) detection, clone P1H12 for MCAM (also known as CD146) detection, and clone DECMA-1 for E-cadherin (also known as CDH1) detection) to the wells / tubes at the predetermined optimal concentration. Incubate the cells with the conjugated antibodies in the dark at 4°C for 15 minutes. After incubation, wash the cells twice with 200 μl (plate) or 2 mL (tube) of cold flow buffer. The washed cells were incubated with an appropriate live / dead distinguishing agent to exclude dead cells from subsequent analyses. The cells were then washed twice with ice-cold flow buffer, resuspended in 0.5 mL of flow buffer, and read immediately on a cytometer.

[0106] Methods / Detection of SRC TGFβ2 secretionTGFβ2 secretion in SRCs was measured using either enzyme-linked immunosorbent assay (ELISA) or the Intelliflex / Luminex assay. In short, frozen SRCs were thawed and seeded in 6-well tissue culture plates at a density of 500,000 cells / well. Twenty-four hours after seeding, the medium was replaced with fresh kidney cell growth medium plus 10 ng PMA (InSolution Phorbol-12-myristate-13-acetate, EMD Millipore, 5.00582.0001) for up to three days. Conditioned medium (supernatant) was collected on days 1, 2, and 3 and stored at -80°C until the TGFβ2 ELISA or Intelliflex / Luminex assay was performed; in the initial assay, the supernatant was collected on day 3 for ELISA. TGFβ2 ELISA was performed according to the manufacturer's instructions for the Quantikine ELISA Human TGF-β2 Kit (R&D Systems, DB-250). The TGFβ2 Intelliflex / Luminex assay was performed using the Luminex TGFβ Triple Kit (R&D Systems) and read on an xMAP Intelliflex reader. Culture medium samples alone were used as negative controls.

[0107] Results / Expression of SRC cell adhesion markers SRC showed upregulation (Log2FC) of cell-cell adhesion genes icam5 (2.5), mcam (1.5), cntnap1 (1.65), col11a1 (3.1), and mmp2 (2.1) and expression of PECAM1 and CDH1 (see example). Figure 1 (and Table 1). Gene ontology confirms the interactionome containing these cell-cell adhesion markers (see, for example, Table 1). Figure 2 ).

[0108] Table 1: Expression of cell adhesion markers in SRC

[0109]

[0110] Results / SRC TGFβ2 Expression: In bulk miR analysis, when SRC was compared with its source renal biopsy tissue, downregulation of hsa-miR-145-5p (log2FC=-6.9) and hsa-miR-199a-5p (log2FC=-5.7) was found in SRC (p < 0.05). adj<0.01). Gene ontology reveals that these miRNAs downregulate the expression of tgfb2 and renal epithelial markers (see, for example...). Figure 3 Consistent with this observation, transcriptome analysis confirmed the upregulation of tgfb2 expression in SRC (log2FC=2.5, p). adj <0.01), included in single-cell analyses (see, for example, Table 2 and...). Figure 4 In ELISA, SRC was also found to secrete TGFβ2 (). Figure 5A n=3; p<0.01 relative to culture medium alone; TGFβ2, ranging from 300 pg / mL to 2000 pg / mL. SRCTGFβ2 secretion is time-dependent (see example). Figure 5B (See Table 3). TGFβ2 secretion in SRCs was confirmed by the Intelliflex / Luminex assay, which is more sensitive than ELISA (see, for example, Table 4). Gene ontology revealed that tgfb2 forms an interaction group with ret, fgf8, lhx1, osr1, and six2 (markers of ureteral buds and cap mesenchyme expressed by SRCs) (see, for example, Table 4). Figure 6 (and Table 2), and GO BP analysis showed that this interaction group was associated with epithelialization of the filtrate components.

[0111] Table 2: Expression of TGFβ2, ureteral bud and cap mesenchymal markers in SRC

[0112]

[0113] Table 3: TGFβ2 secretion in SRC (ELISA)

[0114]

[0115] Table 4: TGFβ2 secretion in SRC (Intelliflex / Luminex)

[0116]

[0117] To confirm the tubule-promoting activity of SRCs, SRCs were placed in culture medium ± hydrogel and stained with epithelial markers GGT and SCL12A1. SRCs assembled into tubules (see example...). Figure 7A ), and found that for GGT1 (see example Figure 7B ) and SCL12A1 (see example) Figure 7C Both can be stained positively, for example, expressing both GGT1 and SCL12A1.

[0118] Further analysis of SRC cell adhesion marker expression resultsFurther SRC cell adhesion marker expression analysis was performed to confirm earlier studies. Additional analysis was performed using SRCs, scRNA-seq analysis, prepared from tissues obtained from the National Disease Research Interchange (NDRI) or from renal biopsies of patients with type 2 diabetes (T2D) and chronic kidney disease (CKD) enrolled in one of two phase 2 clinical trials (002 (NCT02836574) and 007 (NCT05018416)), for example, a phase 2 clinical trial investigating SRC as a potential cell therapy agent. Table 5 provides scRNA-seq expression data for cell adhesion markers.

[0119] Table 5: Expression of cell adhesion markers in SRC (scRNAseq)

[0120]

[0121] 1 The percentage of expression for each gene is calculated as follows: the number of expressed SRCs in the total number of SRCs in the aggregates of 5 patient SRC samples (total number of cells in the 5 aggregates of patient samples = 28,555).

[0122] 2 The percentage of expression for each gene is calculated as follows: the number of expressive SRCs in the total number of SRCs in the aggregates of 17 patient SRC samples (total number of cells in the 17 aggregate patient samples = 204,216).

[0123] The expression of several SRC surface proteins, markers of cell adhesion, was also analyzed by flow cytometry. PECAM, also known as CD31, was detected on the surface of approximately 0.01% to approximately 0.24% of SRCs. ICAM, also known as CD54 (see example...), was detected on the surface of approximately 100% of SRCs. Figure 8 MCAM, also known as CD146, was detected on approximately 99.3% of the SRC surface. (See example) Figure 9 E-cadherin, also known as CDH1, was detected on the surface of approximately 0.1% of SRC (see example). Figure 10 Table 6 below provides a summary of the surface expression data of the above adhesion markers from three SRC samples (PK018, PK019 and HK025).

[0124] Table 6. SRC cell adhesion molecules (positive %)

[0125]

[0126] In summary, the SRC gene expression data presented in this article elucidate the mechanisms by which SRCs mediate regeneration and recovery activity in diseased kidneys. Expression of ureteral bud markers (ret and fgf8) and cap mesenchymal markers (six2, osr1 and lhx1, ret) has identified SRCs as cells potentially capable of reenacting events associated with embryonic kidney development. Identification of SRC TGFβ2 and cell adhesion marker expression suggests that one of the potential therapeutic activities of SRCs is driving tubule formation or the ability to form tubules, which may lead to improvements in electrolyte balance, fluid homeostasis, reabsorption of essential nutrients, urine concentration, and cysteine ​​protease inhibitor C (Cystatin C) metabolism in the context of kidney disease.

[0127] While this disclosure has been specifically shown and described with reference to specific embodiments, some of which are preferred embodiments, those skilled in the art will understand that various changes in form and detail may be made therein without departing from the spirit and scope of this disclosure as disclosed herein. Therefore, it is not intended to be limited to the exact summary and disclosure presented herein.

Claims

1. Methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, including: The enriched heterogeneous renal cell population was assessed to determine whether the cells expressed at least one cell adhesion marker, wherein the at least one cell adhesion marker comprised one or more of the following: intercellular adhesion molecule 5 (ICAM5), melanoma cell adhesion molecule (MCAM), contactin-associated protein 1 (CNTNAP1), or matrix metallopeptidase 2 (MMP2); and If the enriched heterogeneous kidney cell population is found to express at least one of the cell adhesion markers by cytological assays, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

2. The method of claim 1, wherein the determination step comprises determining the percentage of cells expressing the at least one cell adhesion marker in the enriched heterogeneous kidney cell population.

3. The method according to claim 1, wherein the at least one cell adhesion marker includes ICAM5.

4. The method of claim 2, wherein the at least one cell adhesion marker comprises ICAM5, and If more than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population is found to express ICAM5, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

5. The method of claim 2, wherein the at least one cell adhesion marker comprises ICAM5, and If more than about 60% of the cells in the enriched heterogeneous kidney cell population are found to express ICAM5, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

6. The method of claim 1, wherein the at least one cell adhesion marker comprises MCAM.

7. The method of claim 2, wherein the at least one cell adhesion marker comprises MCAM, and If more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population is found to express MCAM, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

8. The method of claim 2, wherein the at least one cell adhesion marker comprises MCAM, and If more than about 65% of the cells in the enriched heterogeneous kidney cell population are found to express MCAM, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

9. The method of claim 1, wherein the at least one cell adhesion marker comprises CNTNAP1.

10. The method of claim 2, wherein the at least one cell adhesion marker comprises CNTNAP1, and If more than about 4% and at most about 15% of the enriched heterogeneous kidney cell population is found to express CNTNAP1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

11. The method of claim 2, wherein the at least one cell adhesion marker comprises CNTNAP1, and If more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population is found to express CNTNAP1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

12. The method of claim 1, wherein the at least one cell adhesion marker comprises MMP2.

13. The method of claim 2, wherein the at least one cell adhesion marker comprises MMP2, and If more than about 8% and at most about 25% of the enriched heterogeneous kidney cell population is found to express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

14. The method of claim 2, wherein the at least one cell adhesion marker comprises MMP2, and If more than about 10% and at most about 35% of the enriched heterogeneous kidney cell population is found to express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

15. The method of claim 2, wherein the at least one cell adhesion marker comprises MMP2, and If more than about 15% and at most about 35% of the enriched heterogeneous kidney cell population is found to express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

16. The method of claim 1, wherein the at least one cell adhesion marker comprises: (a) ICAM5 and MCAM; or (b) ICAM5 and CNTNAP1; or (c) ICAM5 and MMP2; or (d) MCAM and CNTNAP1; or (e) MCAM and MMP2; or (f)CNTNAP1 and MMP2.

17. The method of claim 2, wherein the at least one cell adhesion marker comprises: (a) ICAM5 and MCAM, and If more than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population is found to express ICAM5, and more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population expresses MCAM, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) ICAM5 and CNTNAP1, and If more than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population express ICAM5, and more than about 4% and at most about 15% of the enriched heterogeneous kidney cell population express CNTNAP1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) ICAM5 and MMP2, and If it is determined that more than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population expresses ICAM5, and more than about 8% and at most about 25% of the enriched heterogeneous kidney cell population expresses MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) MCAM and CNTNAP, and If more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population is found to express MCAM, and more than about 4% and at most about 15% of the enriched heterogeneous kidney cell population expresses CNTNAP, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (e) MCAM and MMP2, and If more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population is found to express MCAM, and more than about 8% and at most about 25% of the enriched heterogeneous kidney cell population expresses MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (f) CNTNAP1 and MMP2, and If more than about 4% and up to about 15% of the enriched heterogeneous kidney cell population is found to express CNTNAP, and more than about 8% and up to about 25% of the enriched heterogeneous kidney cell population is found to express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

18. The method of claim 2, wherein the at least one cell adhesion marker comprises: (a) ICAM5 and MCAM, and If more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, and more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) ICAM5 and CNTNAP1, and If more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, and more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) ICAM5 and MMP2, and If more than about 60% of the cells in the enriched heterogeneous kidney cell population are found to express ICAM5, and more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) MCAM and CNTNAP, and If more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, and more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (e) MCAM and MMP2, and If more than about 65% of the cells in the enriched heterogeneous kidney cell population are found to express MCAM, and more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (f) CNTNAP1 and MMP2, and If more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population is found to express CNTNAP1, and more than about 10% and at most about 35% of the enriched heterogeneous kidney cell population is found to express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

19. The method of claim 1, wherein the at least one cell adhesion marker comprises: (a) ICAM5, MCAM and CNTNAP1; or (b) ICAM5, MCAM, and MMP2; or (c) ICAM5, CNTNAP1, and MMP2; or (d)MCAM, CNTNAP1 and MMP2.

20. The method of claim 2, wherein the at least one cell adhesion marker comprises: (a) ICAM5, MCAM and CNTNAP1, and If, in particular, greater than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population are found to express ICAM5, greater than about 15% and at most about 30% of the enriched heterogeneous kidney cell population are found to express MCAM, and greater than about 4% and at most about 15% of the enriched heterogeneous kidney cell population are found to express CNTNAP1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) ICAM5, MCAM and MMP2, and If it is determined that greater than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population expresses ICAM5, greater than about 15% and at most about 30% of the enriched heterogeneous kidney cell population expresses MCAM, and greater than about 8% and at most about 25% of the enriched heterogeneous kidney cell population expresses MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) ICAM5, CNTNAP1 and MMP2, and If, in particular, greater than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population are found to express ICAM5, greater than about 4% and at most about 15% of the enriched heterogeneous kidney cell population are found to express CNTNAP1, and greater than about 8% and at most about 25% of the enriched heterogeneous kidney cell population are found to express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) MCAM, CNTNAP1, and MMP2, and If more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population expresses MCAM, more than about 4% and at most about 15% of the enriched heterogeneous kidney cell population expresses CNTNAP, and more than about 8% and at most about 25% of the enriched heterogeneous kidney cell population expresses MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

21. The method of claim 2, wherein the at least one cell adhesion marker comprises: (a) ICAM5, MCAM and CNTNAP1, and If more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, and more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) ICAM5, MCAM and MMP2, and If it is determined that more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, and more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) ICAM5, CNTNAP1 and MMP2, and If more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, and more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) MCAM, CNTNAP1, and MMP2, and If more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, and more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

22. The method of claim 1, wherein the at least one cell adhesion marker comprises ICAM5, MCAM, CNTNAP1, and MMP2.

23. The method of claim 2, wherein the at least one cell adhesion marker comprises ICAM5, MCAM, CNTNAP1, and MMP2, and If more than 0% and at most about 0.5% of the enriched heterogeneous kidney cell population express ICAM5, more than about 15% and at most about 30% of the enriched heterogeneous kidney cell population express MCAM, more than about 4% and at most about 15% of the enriched heterogeneous kidney cell population express CNTNAP1, and more than about 8% and at most about 25% of the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

24. The method of claim 2, wherein the at least one cell adhesion marker comprises ICAM5, MCAM, CNTNAP1, and MMP2, and If more than about 60% of the cells in the enriched heterogeneous kidney cell population express ICAM5, more than about 65% of the cells in the enriched heterogeneous kidney cell population express MCAM, more than about 15% and at most about 30% of the cells in the enriched heterogeneous kidney cell population express CNTNAP1, and more than about 10% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express MMP2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

25. The method according to any one of the preceding claims, further comprising: The study determined whether the enriched heterogeneous renal cell population expressed at least one renal biomarker. If further analysis reveals that the enriched heterogeneous renal cell population expresses at least one of the aforementioned renal biomarkers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. The at least one renal biomarker includes one or more of the following: sine oculis homeobox homolog 2 (SIX2), transfection rearrangement (RET), odd skip-related 1 (OSR1), lim homeobox 1 (LHX1), or fibroblast growth factor 8 (FGF8).

26. The method of claim 25, wherein determining whether cells in the enriched heterogeneous kidney cell population express the at least one kidney-derived marker comprises determining the percentage of cells in the enriched heterogeneous kidney cell population that express the at least one kidney-derived marker.

27. The method of claim 25, wherein the at least one renal biomarker comprises SIX2.

28. The method of claim 26, wherein the at least one renal biomarker comprises SIX2, and If, in further determination, more than 0% and at most about 6.0% of the enriched heterogeneous kidney cell population express SIX2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

29. The method of claim 25, wherein the at least one renal biomarker comprises OSR1.

30. The method of claim 26, wherein the at least one renal biomarker comprises OSR1, and If it is further determined that more than 0% and at most about 85% of the enriched heterogeneous kidney cell population expresses OSR1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

31. The method of claim 25, wherein the at least one renal biomarker comprises LHX1.

32. The method of claim 26, wherein the at least one renal biomarker comprises LHX1, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

33. The method of claim 25, wherein the at least one renal biomarker comprises RET.

34. The method of claim 26, wherein the at least one renal biomarker comprises RET, and If it is further determined that more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

35. The method of claim 25, wherein the at least one renal biomarker comprises FGF8.

36. The method of claim 26, wherein the at least one renal biomarker comprises FGF8, and If it is further determined that more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

37. The method of claim 25, wherein the at least one renal biomarker comprises: (a) SIX2 and OSR1; or (b) SIX2 and LHX1; or (c) SIX2 and RET; or (d) SIX2 and FGF8; or (e)OSR1 and LHX1; or (f) OSR1 and RET; or (g)OSR1 and FGF8; or (h)LHX1 and RET; or (i) LHX1 and FGF8; or (j)RET and FGF8.

38. The method of claim 26, wherein the at least one renal biomarker comprises: (a) SIX2 and OSR1, and If, further determination reveals that more than 0% and at most approximately 6% of the enriched heterogeneous kidney cell population expresses SIX2, and more than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) SIX2 and LHX1, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, and more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) SIX2 and RET, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, and more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) SIX2 and FGF8, and If, further determination reveals that more than 0% and at most approximately 6% of the enriched heterogeneous kidney cell population expresses SIX2, and more than 0% and at most approximately 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (e)OSR1 and LHX1, and If, upon further determination, greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, and greater than approximately 8% and at most approximately 58% of the enriched heterogeneous kidney cell population expresses LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (f) OSR1 and RET, and If, further determination reveals that greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, and greater than 0% and at most approximately 90% of the enriched heterogeneous kidney cell population expresses RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (g)OSR1 and FGF8, and If, further determination reveals that greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, and greater than 0% and at most approximately 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (h)LHX1 and RET, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (i) LHX1 and FGF8, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (j)RET and FGF8, and If it is further determined that more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

39. The method of claim 25, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1, and LHX1; or (b) SIX2, OSR1, and RET; or (c) SIX2, OSR1, and FGF8; or (d) SIX2, LHX1, and RET; or (e) SIX2, LHX1, and FGF8; or (f) SIX2, RET, and FGF8; or (g) OSR1, LHX1, and RET; or (h)OSR1, LHX1, and FGF8; or (i) OSR1, RET, and FGF8; or (j)LHX1, RET and FGF8.

40. The method of claim 26, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1 and LHX1, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than about 0% and at most about 85% of the enriched heterogeneous kidney cell population express OSR1, and more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) SIX2, OSR1 and RET, and If, upon further determination, more than 0% and at most approximately 6% of the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population express OSR1, and more than 0% and at most approximately 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) SIX2, OSR1 and FGF8, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than about 0% and at most about 85% of the enriched heterogeneous kidney cell population express OSR1, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) SIX2, LHX1, and RET, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (e)SIX2, LHX1 and FGF8, and If, upon further determination, more than 0% and at most about 6.0% of the enriched heterogeneous kidney cell population expresses SIX2, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population expresses LHX1, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (f) SIX2, RET and FGF8, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than about 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (g) OSR1, LHX1, and RET, and If, upon further determination, greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, greater than approximately 8% and at most approximately 58% of the enriched heterogeneous kidney cell population expresses LHX1, and greater than 0% and at most approximately 90% of the enriched heterogeneous kidney cell population expresses RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (h)OSR1, LHX1 and FGF8, and If, upon further determination, greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, greater than approximately 8% and at most approximately 58% of the enriched heterogeneous kidney cell population expresses LHX1, and greater than 0% and at most approximately 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (i) OSR1, RET and FGF8, and If, further determination is made that greater than 0% and at most about 85% of the enriched heterogeneous kidney cell population expresses OSR1, greater than about 0% and at most about 90% of the enriched heterogeneous kidney cell population expresses RET, and greater than 0% and at most about 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (j)LHX1, RET and FGF8, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

41. The method of claim 25, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1, LHX1, and RET; or (b) SIX2, OSR1, LHX1, and FGF8; or (c) SIX2, LHX1, RET, and FGF8; or (d) SIX2, OSR1, RET, and FGF8; or (e)OSR1, LHX1, RET and FGF8.

42. The method of claim 26, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1, LHX1 and RET, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the cells in the enriched heterogeneous kidney cell population express OSR1, more than about 8% and at most about 58% of the cells in the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (b) SIX2, OSR1, LHX1 and FGF8, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the cells in the enriched heterogeneous kidney cell population express OSR1, more than about 8% and at most about 58% of the cells in the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 59% of the cells in the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (c) SIX2, LHX1, RET and FGF8, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than about 8% and at most about 58% of the cells in the enriched heterogeneous kidney cell population express LHX1, more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the cells in the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (d) SIX2, OSR1, RET and FGF8, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the cells in the enriched heterogeneous kidney cell population express OSR1, more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the cells in the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (e) OSR1, LHX1, RET and FGF8, and If, further determination is made that more than 0% and at most about 85% of the enriched heterogeneous kidney cell population expresses OSR1, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population expresses LHX1, more than 0% and at most about 90% of the enriched heterogeneous kidney cell population expresses RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

43. The method of claim 25, wherein the at least one renal biomarker comprises SIX2, OSR1, LHX1, RET, and FGF8.

44. The method of claim 26, wherein the at least one renal biomarker comprises SIX2, OSR1, LHX1, RET, and FGF, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the enriched heterogeneous kidney cell population express OSR1, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

45. The method according to any one of the preceding claims, further comprising: The study determined whether the enriched heterogeneous renal cell population expressed at least one additional cell adhesion marker. If, further analysis reveals that the enriched heterogeneous kidney cell population expresses at least one additional cell adhesion marker, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. The at least one additional cell adhesion marker mentioned above includes: type XI collagen α1 (COL11A1), platelet endothelial cell adhesion molecule (PECAM) 1, or E-cadherin.

46. ​​The method of claim 45, wherein determining whether cells in the enriched heterogeneous kidney cell population express the at least one additional cell adhesion marker comprises determining the percentage of cells in the enriched heterogeneous kidney cell population that express the at least one additional cell adhesion marker.

47. The method of claim 46, wherein the at least one additional cell adhesion marker comprises COL11A1, and If, upon further determination, more than about 20% and at most about 55% of the cells in the enriched heterogeneous kidney cell population express COL11A1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

48. The method of claim 46, wherein the at least one additional cell adhesion marker comprises COL11A1, and If, upon further determination, more than about 20% and at most about 40% of the cells in the enriched heterogeneous kidney cell population express COL11A1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

49. The method of claim 46, wherein the at least one additional cell adhesion marker comprises COL11A1, and If, upon further determination, more than about 25% and at most about 55% of the cells in the enriched heterogeneous kidney cell population express COL11A1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

50. The method according to any one of claims 46-49, wherein the at least one additional cell adhesion marker comprises PECAM1, and If, upon further determination, more than about 0.1% and at most about 0.4% of the enriched heterogeneous kidney cell population expresses PECAM1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

51. The method according to any one of claims 46-50, wherein the at least one additional cell adhesion marker comprises E-cadherin, and If, upon further determination, more than about 8% and at most about 35% of the enriched heterogeneous kidney cell population express E-cadherin, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

52. The method according to any one of claims 46-50, wherein the at least one additional cell adhesion marker comprises E-cadherin, and If, upon further determination, more than about 15% and at most about 35% of the cells in the enriched heterogeneous kidney cell population express E-cadherin, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

53. The method according to any one of claims 46-50, wherein the at least one additional cell adhesion marker comprises E-cadherin, and If it is further determined that more than 0% and at most about 5% of the cells in the enriched heterogeneous kidney cell population express E-cadherin, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

54. The method according to any one of the preceding claims, further comprising: The study determined whether the enriched heterogeneous kidney cell population expressed transforming growth factor β2 (TGFβ2). If the enriched heterogeneous kidney cell population is further found to express TGFβ2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

55. The method of claim 54, wherein determining whether the cells in the enriched heterogeneous kidney cell population express TGFβ2 comprises determining the percentage of cells in the enriched heterogeneous kidney cell population that express TGFβ2.

56. The method of claim 55, wherein if it is further determined that more than about 50% of the cells in the enriched heterogeneous kidney cell population express TGFβ2, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

57. The method of claim 54, wherein determining whether the cells of the enriched heterogeneous kidney cell population express TGFβ2 comprises determining whether the cells of the enriched heterogeneous kidney cell population secrete TGFβ2.

58. The method of claim 57, wherein if it is further determined that each 1,000,000 enriched heterogeneous kidney cell population secretes at least about 1.0 ng of TGFβ2, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

59. The method of claim 57, wherein if it is further determined that each 1,000,000 enriched heterogeneous kidney cell population secretes at least about 2.0 ng of TGFβ2, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

60. The method of claim 57, wherein the enriched heterogeneous kidney cell population is identified as having therapeutic potential if it is further determined that each 1,000,000 enriched heterogeneous kidney cell population secretes at least about 2.4 ng of TGFβ2.

61. The method of claim 57, wherein if it is further determined that each 1,000,000 enriched heterogeneous kidney cell population secretes about 3.0 ng to about 20 ng of TGFβ2, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

62. The method according to any one of the preceding claims, further comprising: The enriched heterogeneous kidney cell population was determined to express at least one further biomarker. If the enriched heterogeneous kidney cell population is further found to express at least one of the further markers, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. The at least one further marker mentioned above includes nephrin, podocin, or a receptor that activates C kinase (RACK-1).

63. The method of claim 62, wherein determining whether cells in the enriched heterogeneous kidney cell population express the at least one further marker comprises determining the percentage of cells in the enriched heterogeneous kidney cell population expressing the at least one further marker.

64. The method of claim 63, wherein the at least one further biomarker comprises nephrotic protein, and If, upon further determination, more than about 4% and up to about 99% of the cells in the heterogeneous kidney cell population express nephrotic proteins, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

65. The method of claim 63, wherein the at least one further marker comprises a foot spike protein, and If it is further determined that more than 90% of the cells in the heterologous kidney cell population express podocyte protein, then the enriched heterologous kidney cell population is identified as having therapeutic potential.

66. The method of claim 63, wherein the at least one further marker comprises RACK-1, and If, upon further determination, at least approximately 85% of the cells in the heterogeneous kidney cell population express RACK-1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

67. Methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, including: The expression of TGFβ2 in the enriched heterogeneous kidney cell population was measured; and If it is determined that: (i) More than 50% of the cells in the enriched heterogeneous kidney cell population express TGFβ2; and / or (ii) TGFβ2 is secreted by cells from the enriched heterogeneous kidney cell population at a rate of at least about 1.0 ng / 1,000,000 enriched heterogeneous kidney cell populations. The enriched heterogeneous renal cell population was then identified as having therapeutic potential.

68. The method of claim 67, wherein if (i) more than about 50% of the cells in the enriched heterogeneous kidney cell population are found to express TGFβ2, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

69. The method of claim 68, wherein if about 50% to about 80% of the enriched heterogeneous kidney cell population is found to express TGFβ2, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

70. The method of claim 67, wherein if (ii) TGFβ2 is determined to be secreted by cells of the enriched heterogeneous kidney cell population at an amount of at least about 1.0 ng / 1,000,000 enriched heterogeneous kidney cell population, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

71. The method of claim 70, wherein the enriched heterogeneous kidney cell population is identified as having therapeutic potential if TGFβ2 is determined to be secreted by cells of the enriched heterogeneous kidney cell population at an amount of at least about 2.0 ng / 1,000,000 enriched heterogeneous kidney cell population cells.

72. The method of claim 71, wherein the enriched heterogeneous kidney cell population is identified as having therapeutic potential if TGFβ2 is determined to be secreted by cells of the enriched heterogeneous kidney cell population at an amount of at least about 2.4 ng / 1,000,000 enriched heterogeneous kidney cell population cells.

73. The method of claim 72, wherein the enriched heterogeneous kidney cell population is identified as having therapeutic potential if TGFβ2 is determined to be secreted by cells of the enriched heterogeneous kidney cell population at an amount of at least about 3.0 ng / 1,000,000 enriched heterogeneous kidney cell population cells.

74. The method of claim 73, wherein the enriched heterogeneous kidney cell population is identified as having therapeutic potential if TGFβ2 is determined to be secreted by cells of the enriched heterogeneous kidney cell population in an amount of about 3.0 ng to about 20 ng per 1,000,000 enriched heterogeneous kidney cell population cells.

75. The method of claim 67, wherein if (ii) TGFβ2 is determined to be secreted by cells of the enriched heterogeneous kidney cell population at an amount of at least about 0.7 ng / 1,000,000 enriched heterogeneous kidney cell population over a period of about 24 hours, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

76. The method of claim 75, wherein the amount is at least about 2.0 ng / 1,000,000 enriched heterogeneous kidney cell populations over a period of about 24 hours.

77. The method of claim 67, wherein if (ii) TGFβ2 is determined to be secreted by the cells of the enriched heterogeneous kidney cell population at a rate of at least about 3.5 ng / 1,000,000 enriched heterogeneous kidney cell population over a period of about 48 hours, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

78. The method of claim 67, wherein if (ii) TGFβ2 is determined to be secreted by the cells of the enriched heterogeneous kidney cell population at an amount of at least about 8.0 ng / 1,000,000 enriched heterogeneous kidney cell population over a period of about 72 hours, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

79. The method according to claim 68 or 69, wherein if (ii) TGFβ2 is further determined to be secreted by cells of the enriched heterogeneous kidney cell population in an amount of at least about 1.0 ng / 1,000,000 enriched heterogeneous kidney cell population, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

80. The method according to claim 68 or 69, wherein if (ii) TGFβ2 is further determined to be secreted by the cells of the enriched heterogeneous kidney cell population in an amount of at least about 2.0 ng / 1,000,000 enriched heterogeneous kidney cell population, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

81. The method according to claim 68 or 69, wherein the enriched heterogeneous kidney cell population is identified as having therapeutic potential if it is further determined that TGFβ2 is secreted by cells of the enriched heterogeneous kidney cell population in an amount of at least about 3.0 ng / 1,000,000 enriched heterogeneous kidney cell population cells.

82. The method according to claim 68 or 69, wherein if TGFβ2 is further determined to be secreted by cells of the enriched heterogeneous kidney cell population in an amount of about 3.0 ng to about 20 ng / 1,000,000 enriched heterogeneous kidney cell population, the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

83. The method according to claim 65 or 69, wherein the enriched heterogeneous kidney cell population is identified as having therapeutic potential if it is further determined that TGFβ2 is secreted by the cells of the enriched heterogeneous kidney cell population in the following amounts: Within a period of approximately 24 hours, at least approximately 0.7 ng / 1,000,000 enriched heterogeneous kidney cell populations were observed. Within a timeframe of approximately 48 hours, at least approximately 3.5 ng / 1,000,000 enriched heterogeneous renal cell populations were observed; and / or Over a period of approximately 72 hours, at least approximately 8.0 ng / 1,000,000 enriched heterogeneous kidney cell populations were observed.

84. The method of claim 83, wherein, over a period of approximately 24 hours, the amount of TGFβ2 secreted by the cells of the enriched heterogeneous kidney cell population is at least approximately 2.0 ng / 1,000,000 enriched heterogeneous kidney cell population cells.

85. The method of claim 84, wherein, over a period of approximately 24 hours, the amount of TGFβ2 secreted by the cells of the enriched heterogeneous kidney cell population is at least approximately 2.5 ng / 1,000,000 enriched heterogeneous kidney cell population cells.

86. The method according to any one of claims 67-85, further comprising determining whether the cells of the enriched heterogeneous kidney cell population express: At least one renal biomarker, If further analysis reveals that the enriched heterogeneous renal cell population expresses at least one of the aforementioned renal biomarkers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. The at least one renal biomarker includes one or more of the following: SIX2, RET, OSR1, LHX1, or FGF8.

87. The method of claim 86, wherein determining whether cells in the enriched heterogeneous kidney cell population express the at least one kidney-derived marker comprises determining the percentage of cells in the enriched heterogeneous kidney cell population that express the at least one kidney-derived marker.

88. The method of claim 86, wherein the at least one renal biomarker comprises SIX2.

89. The method of claim 87, wherein the at least one renal biomarker comprises SIX2, and If, upon further determination, more than 0% and at most about 6.0% of the enriched heterogeneous kidney cell population express SIX2, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

90. The method of claim 86, wherein the at least one renal biomarker comprises OSR1.

91. The method of claim 87, wherein the at least one renal biomarker comprises OSR1, and If it is further determined that more than 0% and at most about 85% of the enriched heterogeneous kidney cell population expresses OSR1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

92. The method of claim 86, wherein the at least one renal biomarker comprises LHX1.

93. The method of claim 87, wherein the at least one renal biomarker comprises LHX1, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

94. The method of claim 86, wherein the at least one renal biomarker comprises RET.

95. The method of claim 87, wherein the at least one renal biomarker comprises RET, and If it is further determined that more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

96. The method of claim 86, wherein the at least one renal biomarker comprises FGF8.

97. The method of claim 87, wherein the at least one renal biomarker comprises FGF8, and If it is further determined that more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

98. The method of claim 86, wherein the at least one renal biomarker comprises: (a) SIX2 and OSR1; or (b) SIX2 and LHX1; or (c) SIX2 and RET; or (d) SIX2 and FGF8; or (e)OSR1 and LHX1; or (f) OSR1 and RET; or (g)OSR1 and FGF8; or (h)LHX1 and RET; or (i) LHX1 and FGF8; or (j)RET and FGF8.

99. The method of claim 87, wherein the at least one renal biomarker comprises: (a) SIX2 and OSR1, and If, further determination reveals that more than 0% and at most approximately 6% of the enriched heterogeneous kidney cell population expresses SIX2, and more than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) SIX2 and LHX1, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, and more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) SIX2 and RET, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, and more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) SIX2 and FGF8, and If, further determination reveals that more than 0% and at most approximately 6% of the enriched heterogeneous kidney cell population expresses SIX2, and more than 0% and at most approximately 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (e)OSR1 and LHX1, and If, upon further determination, greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, and greater than approximately 8% and at most approximately 58% of the enriched heterogeneous kidney cell population expresses LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (f) OSR1 and RET, and If, further determination reveals that greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, and greater than 0% and at most approximately 90% of the enriched heterogeneous kidney cell population expresses RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (g)OSR1 and FGF8, and If, further determination reveals that greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, and greater than 0% and at most approximately 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (h)LHX1 and RET, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (i) LHX1 and FGF8, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (j)RET and FGF8, and If it is further determined that more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

100. The method of claim 86, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1, and LHX1; or (b) SIX2, OSR1, and RET; or (c) SIX2, OSR1, and FGF8; or (d) SIX2, LHX1, and RET; or (e) SIX2, LHX1, and FGF8; or (f) SIX2, RET, and FGF8; or (g) OSR1, LHX1, and RET; or (h)OSR1, LHX1, and FGF8; or (i) OSR1, RET, and FGF8; or (j)LHX1, RET and FGF8.

101. The method of claim 87, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1 and LHX1, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than about 0% and at most about 85% of the enriched heterogeneous kidney cell population express OSR1, and more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (b) SIX2, OSR1 and RET, and If, upon further determination, more than 0% and at most approximately 6% of the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population express OSR1, and more than 0% and at most approximately 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (c) SIX2, OSR1 and FGF8, and If, further determination reveals that more than 0% and at most about 6% of the enriched heterogeneous kidney cell population expresses SIX2, more than about 0% and at most about 85% of the enriched heterogeneous kidney cell population expresses OSR1, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (d) SIX2, LHX1, and RET, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (e)SIX2, LHX1 and FGF8, and If, upon further determination, more than 0% and at most about 6.0% of the enriched heterogeneous kidney cell population expresses SIX2, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population expresses LHX1, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (f) SIX2, RET and FGF8, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than about 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (g) OSR1, LHX1, and RET, and If, upon further determination, greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, greater than approximately 8% and at most approximately 58% of the enriched heterogeneous kidney cell population expresses LHX1, and greater than 0% and at most approximately 90% of the enriched heterogeneous kidney cell population expresses RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (h)OSR1, LHX1 and FGF8, and If, upon further determination, greater than 0% and at most approximately 85% of the enriched heterogeneous kidney cell population expresses OSR1, greater than approximately 8% and at most approximately 58% of the enriched heterogeneous kidney cell population expresses LHX1, and greater than 0% and at most approximately 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (i) OSR1, RET and FGF8, and If, further determination shows that more than 0% and at most about 85% of the enriched heterogeneous kidney cell population expresses OSR1, more than about 0% and at most about 90% of the enriched heterogeneous kidney cell population expresses RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential; or (j)LHX1, RET and FGF8, and If, upon further determination, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

102. The method of claim 86, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1, LHX1, and RET; or (b) SIX2, OSR1, LHX1, and FGF8; or (c) SIX2, LHX1, RET, and FGF8; or (d) SIX2, OSR1, RET, and FGF8; or (e)OSR1, LHX1, RET and FGF8.

103. The method of claim 87, wherein the at least one renal biomarker comprises: (a) SIX2, OSR1, LHX1 and RET, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the cells in the enriched heterogeneous kidney cell population express OSR1, more than about 8% and at most about 58% of the cells in the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (b) SIX2, OSR1, LHX1 and FGF8, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the cells in the enriched heterogeneous kidney cell population express OSR1, more than about 8% and at most about 58% of the cells in the enriched heterogeneous kidney cell population express LHX1, and more than 0% and at most about 59% of the cells in the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (c) SIX2, LHX1, RET and FGF8, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than about 8% and at most about 58% of the cells in the enriched heterogeneous kidney cell population express LHX1, more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the cells in the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (d) SIX2, OSR1, RET and FGF8, and If, further, it is determined that more than 0% and at most about 6% of the cells in the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the cells in the enriched heterogeneous kidney cell population express OSR1, more than 0% and at most about 90% of the cells in the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the cells in the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential. (e) OSR1, LHX1, RET and FGF8, and If, further determination is made that more than 0% and at most about 85% of the enriched heterogeneous kidney cell population expresses OSR1, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population expresses LHX1, more than 0% and at most about 90% of the enriched heterogeneous kidney cell population expresses RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population expresses FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

104. The method of claim 86, wherein the at least one renal biomarker comprises SIX2, OSR1, LHX1, RET, and FGF8.

105. The method of claim 87, wherein the at least one renal biomarker comprises SIX2, OSR1, LHX1, RET, and FGF, and If, upon further determination, more than 0% and at most about 6% of the enriched heterogeneous kidney cell population express SIX2, more than 0% and at most about 85% of the enriched heterogeneous kidney cell population express OSR1, more than about 8% and at most about 58% of the enriched heterogeneous kidney cell population express LHX1, more than 0% and at most about 90% of the enriched heterogeneous kidney cell population express RET, and more than 0% and at most about 59% of the enriched heterogeneous kidney cell population express FGF8, then the enriched heterogeneous kidney cell population is identified as having therapeutic potential.

106. The enriched heterogeneous kidney cell population identified by the method according to any one of claims 1-105.

107. A pharmaceutical composition comprising the enriched heterogeneous renal cell population as described in claim 106.

108. A method for treating kidney disease in patients in need, the method comprising: Administer a therapeutically effective amount of the pharmaceutical composition according to claim 107.

109. Use of the pharmaceutical composition according to claim 107 in the preparation of a medicament for treating kidney diseases.

110. The enriched heterogeneous kidney cell population of claim 106, wherein the population is prepared via a method including a density gradient separation step.

111. The enriched heterogeneous kidney cell population of claim 110, wherein the population comprises cells with a buoyancy density greater than about 1.04 g / mL.

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