Selected renal cell population cells, characteristics and uses thereof

Abstract

Methods for identifying enriched heterogeneous renal cell populations with therapeutic potential, the enriched heterogeneous renal cell populations with therapeutic potential, and their uses. In some embodiments, it includes 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 of: (i) ureteral bud and / or cap mesenchymal (UB / CM) markers; and / or (ii) podocyte markers.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 589,937, filed October 12, 2023, and U.S. Provisional Patent Application Serial No. 63 / 652,522, filed May 28, 2024. The disclosure of the above applications, in its entirety (including any drawings), is expressly incorporated herein by reference. Background Technology

[0003] Given the increasing prevalence of diabetes and metabolic syndrome, the incidence and prevalence of chronic kidney disease (CKD) are rising (Thurlow JS, et al., Am J Nephrol. 2021; 52:98-107). Without treatment, CKD can progress to end-stage renal disease (ESRD) and require renal replacement therapy. Dialysis, associated with increased morbidity and mortality, remains not only a bridge to transplantation but also represents a significant burden on federal health finances (Thurlow JS, et al., Am J Nephrol. 2021; 52:98-107; Murphy D, et al., Intern Med 2016; 65:473-481; Hill NR, et al., PLoS One 11:e0158765. doi: 10.1371 / journal.pone.0158765; Glassock RJ, et al., Nat Rev Nephrol. 2017; 13:104-114; Migliori M, et al., J Nephrol. 2017, 30:373-383). Given the often refractory nature of treatments addressing the underlying causes of the disease, there is an urgent need for methods to repair renal microstructure, restore renal function, and delay or avoid the need for dialysis.

[0004] Autologous cell therapy uses cells from an individual, which are typically cultured and expanded in vitro before being reintroduced into the individual. Advantages of this approach include reduced risks of alloimmune events, bioincompatibility, and disease transmission (Kazmi B, et al., Wounds 2009; 9:234-242). Autologous selected kidney cell (SRC) therapy utilizes kidney cells derived from a biopsy, with a selected subset implanted into the donor kidney (Stavas J, et al., KI Reports 2022; 7:1619-1629; Stavas J, et al., Blood Purif.2023; 52:114-121). Phenotypic marker studies have shown that SRC express ureteral buds (UB), cap mesenchymal (CM), and podocyte markers (Stavas J, et al., KI Reports2022; 7:1619-1629), indicating that these cells possess potential for kidney repair and recovery. In the cohort of diabetic nephropathy (DKD) with increased risk of ESRD, individuals were randomly assigned to the SRC (REACT) cohort. ™ This may be related to the stability of renal function (Stavas J, et al., KI Reports 2022; 7:1619-1629; Stavas J, et al., Blood Purif. 2023; 52:114-121).

[0005] There is a need in the art to identify heterogeneous populations of renal cells, such as SRCs, with potential therapeutic activity that could delay or prevent the progression of CKD patients to ESRD, said identification being performed, for example, via renal repair and recovery capabilities. 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: (i) a ureteric bud and / or cap mesenchyme (UB / CM) marker; and / or (ii) a podocyte marker. At least one UB / CM marker comprises one or more of the following: Wingless-related integration site family member 11 (WNT11), Gremlin (GREM)1, E26 transformation-specific variant transcription 5 (ETV5), Homeobox (HOX)11, or neural cell adhesion molecule (NCAM)1. At least one podocyte marker comprises one or more of the following: thrombospondin type 1 domain containing (THSD) 7A, protein tyrosine phosphatase receptor type O (PTPRO), matrix metalloproteinase (MPP) 5, tubulin polymerization promoting protein (TPP) 3, or lim homeobox transcription factor 1 beta (LMX1B). If the enriched heterogeneous renal cell population is found to express at least one of the following: (i) UB / CM marker and / or (ii) podocyte marker, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

[0007] This disclosure describes an additional method for identifying enriched heterogeneous renal cell populations as having therapeutic potential. The percentage of renal cells in the population expressing ureteric bud, cap-mesenchymal, and podocyte characteristic genes is determined. A renal cell population is identified as having therapeutic potential if (i) approximately 20% to approximately 40% of the renal cells in the population express ureteric bud and cap-mesenchymal cell characteristic genes; and (ii) approximately 50% to approximately 65% ​​of the renal cells in the population express podocyte characteristic genes. This population is prepared from non-fetal renal tissue. Attached Figure Description

[0008] Figure 1 : Shows the diameter size of human SRC-derived organoids cultured within 18 days ( m).

[0009] Figures 2A-2D Figure 2A shows images of human SRC-derived organoids stained with immunofluorescence antibodies. Organoid expression of ureteral bud + cap mesenchymal cell markers (Hoxa11 (Fig. 2B), CK8 (Fig. 2C), and Meis2 (Fig. 2D)) was detected. Figure 2A shows organoids stained with isotype-matched antibody controls.

[0010] Figures 3A-3B Images of human SRC-derived organoids stained with immunofluorescence antibodies are shown. Organoid expression of glomerular epithelial markers (Thds7a (Fig. 3A) and Ptpro (Fig. 3B)) was detected.

[0011] Figures 4A-4B Images of human SRC-derived organoids stained with immunofluorescence antibodies are shown. Organoid expression of proximal tubule markers (LTL (Fig. 4A) and CD13 (Fig. 4A and 4B)) was detected.

[0012] Figures 5A-5B Figure 5A shows images of human SRC-derived organoids stained with immunofluorescence antibodies. Organoid expression of the epithelial cell division cycle marker Cdc6 was detected (Figure 5B). Figure 5B shows organoids stained with isotype-matched antibody controls.

[0013] Figures 6A-6B Images of human SRC-derived organoids cultured for 8 days (Fig. 6A) and 17 days (Fig. 6B) after being prepared as sections and stained with periodic acid Schiff (PAS) are shown. On day 8 of culture, structures with a glomerular-like appearance were observed, including parietal epithelial cell (PEC)-like margins (Fig. 6A, arrow). On day 17 of culture, structures exhibiting nephron-like features were observed, including glomerular cluster-like structures (Fig. 6B, arrow).

[0014] Figures 7A-7B Figure 7A shows tubular structural features in images of 17-day-old cultured human SRC-derived organoids prepared as sections and stained with periodic acid Schiff (PAS) (within the rectangle in Figure 7A). Figure 7B is a magnified view of the region inside the rectangle in Figure 7A.

[0015] Figure 8 This study reveals the opn / ssp1-cav1-mmp9-icam1-cxcr4 axis, as revealed by gene ontology, which regulates kidney migration, cell-cell adhesion, and wound healing.

[0016] Figure 9: Shows genes and / or proteins expressed by SRC and cells during kidney development.

[0017] Figure 10 The shared podocyte-PEC and SRC transcriptomes were shown.

[0018] Figure 11 Uniform manifold approximation and projection (UMAP) shows SRC expression of hes1. Dark blue / dark gray dots represent cells expressing hes1. Light gray dots represent cells not expressing hes1. The majority of cells (approximately 90%) in this particular SRC population express hes1. Detailed Implementation

[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 identified as having therapeutic potential.

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

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

[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: (i) UB / CM marker; and / or (ii) podocyte marker. In some cases, at least one marker may be (i) a UB / CM marker. Where at least one marker is at least one UB / CM marker, at least one UB / CM marker may be or include any of the following: Wingless-associated integration site family member 11 (WNT11), Gremlin (GREM)1, E26 transformation-specific (ETS) variant transcription 5 (ETV5), homeobox (HOX)11, or neural cell adhesion molecule (NCAM)1. In some cases, where at least one marker is at least one UB / CM marker, at least one UB / CM marker may be two or more of the following: WNT11, GREM1, ETV5, HOX11, and / or NCAM1. At least two UB / CM markers may be or include: (a) WNT11 and GREM1; (b) WNT11 and ETV5; (c) WNT11 and HOX11; (d) WNT11 and NCAM1; (e) GREM1 and ETV5; (f) GREM1 and HOX11; (g) GREM1 and NCAM1; (h) ETV5 and HOX11; (i) ETV5 and NCAM1; or (j) HOX11 and NCAM1. In other cases, at least one marker is at least one UB / CM marker, and at least one UB / CM marker may be any three or more of the following: WNT11, GREM1, ETV5, HOX11, and / or NCAM1. At least three UB / CM markers may be or include: (a) WNT11, GREM1, and ETV5; (b) WNT11, GREM1, and HOX11; (c) WNT11, GREM1, and NCAM1; (d) WNT11, ETV5, and HOX11; (e) WNT11, ETV5, and NCAM1; (f) WNT11, HOX11, and NCAM1; (g) GREM1, ETV5, and HOX11; (h) GREM1, ETV5, and NCAM1; (i) GREM1, HOX11, and NCAM1; or (j) ETV, HOX11, and NCAM1. In other cases, where at least one marker is at least one UB / CM marker, at least one UB / CM marker may be any four or more of the following: WNT11, GREM1, ETV5, HOX11, and / or NCAM1.At least four UB / CM markers may be or include: (a) WNT11, GREM1, ETV5, and HOX11; (b) WNT11, GREM1, ETV5, and NCAM1; (c) WNT11, ETV5, HOX11, and NCAM1; (d) WNT11, GREM1, HOX11, and NCAM1; or (e) GREM1, ETV5, HOX11, and NCAM1. In other cases, where at least one marker is at least one UB / CM marker, at least one UB / CM marker may be or include WNT11, GREM1, ETV5, HOX11, and / or NCAM1.

[0023] In methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, a method for determining whether the cells of the enriched heterogeneous renal cell population express at least one of the following: (i) UB / CM markers; and / or (ii) podocyte markers, wherein at least one marker may be (ii) a podocyte marker. Where at least one marker is at least one podocyte marker, the at least one podocyte marker may be or include any of the following: platelet-reactive protein type 1 domain protein (THSD) 7A, protein tyrosine phosphatase receptor type O (PTPRO), matrix metalloproteinase (MPP) 5, tubulin polymerization-promoting protein (TPPP) 3, or lim homeobox transcription factor 1β (LMX1B). In some cases, where at least one marker is at least one podocyte marker, the at least one podocyte marker may be two or more of the following: THSD7A, PTPRO, MPP5, TPPP3, and / or LMX1B. At least two or more podocyte markers may be or include: (a) THSD7A and PTPRO; (b) THSD7A and MPP5; (c) THSD7A and TPPP3; (d) THSD7A and LMX1B; (e) PTPRO and MPP5; (f) PTPRO and TPPP3; (g) PTPRO and LMX1B; (h) MPP5 and TPPP3; (i) MPP5 and LMX1B; or (j) TPPP3 and LMX1B. In other cases, where at least one marker is at least one podocyte marker, at least one podocyte marker may be any three or more of the following: THSD7A, PTPRO, MPP5, TPPP3, and / or LMX1B. Three or more podocyte markers may be or include: (a) THSD7A, PTPRO, and MPP5; (b) THSD7A, PTPRO, and TPPP3; (c) THSD7A, PTPRO, and LMX1B; (d) THSD7A, MPP5, and TPPP3; (e) THSD7A, MPP5, and LMX1B; (f) THSD7A, TPPP3, and LMX1B; (g) PTPRO, MPP5, and TPPP3; (h) PTPRO, MPP5, and LMX1B; (i) PTPRO, TPPP3, and LMX1B; or (j) MPP5, TPPP3, and LMX1B. In other cases, where at least one marker is at least one podocyte marker, the at least one podocyte marker may be any four or more of THSD7A, PTPRO, MPP5, TPPP3, and / or LMX1B.Four or more podocyte markers may be or include: (a) THSD7A, PTPRO, MPP5, and TPPP3; (b) THSD7A, PTPRO, MPP5, and LMX1B; (c) THSD7A, MPP5, TPPP3, and LMX1B; (d) THSD7A, PTPRO, TPPP3, and LMX1B; or (e) GREM1, MPP5, TPPP3, and LMX1B. In other cases, where at least one marker is at least one podocyte marker, at least one podocyte marker may be or include THSD7A, PTPRO, MPP5, TPPP3, and / or LMX1B.

[0024] In methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, a method may be used to determine whether the cells of the enriched heterogeneous renal cell population express at least one of the following: (i) UB / CM markers; and / or (ii) podocyte markers, wherein at least one marker may include both (i) one or more UB / CM markers and (ii) one or more podocyte markers. In these methods, at least one UB / CM marker may be or include any one, any combination of two, any combination of three, any combination of four, or all five of WNT11, GREM1, ETV5, HOX11, and / or NCAM1, for example, as described above herein, and at least one podocyte marker may be or include any one, any combination of two, any combination of three, any combination of four, or all five of THSD7A, PTPRO, MPP5, TPPP3, or LMX1B.

[0025] Among these methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, determining that the enriched heterogeneous renal cell population expresses (i) at least one (any one, at least two, at least three, at least four or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte markers can identify the enriched heterogeneous renal cell population as having therapeutic potential.

[0026] In methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, determining the expression of (i) at least one (any one, at least two, at least three, at least four, or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four, or five) podocyte markers may further include determining the percentage of cells in the enriched heterogeneous renal cell population expressing (i) at least one (any one, at least two, at least three, at least four, or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four, or five) podocyte markers. If the percentage of cells expressing (i) at least one (any one, at least two, at least three, at least four or five) UB / CM markers and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte markers in an enriched heterogeneous renal cell population is determined, then the enriched heterogeneous renal cell population can be identified as having therapeutic potential if approximately a certain or specific percentage of cells in the enriched heterogeneous renal cell population express (i) at least one (any one, at least two, at least three, at least four or five) UB / CM markers and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte markers.

[0027] In this method, if the percentage of cells expressing WNT11 in an enriched heterogeneous renal cell population is determined, the enriched heterogeneous renal cell population can be identified as having therapeutic potential when the percentage of cells expressing WNT11 is greater than 0%, greater than 0% and at most about 10%, greater than 0% and at most about 7.5%, greater than 0% and at most about 5%, greater than 0% and at most about 2.5%, greater than 0% and at most about 2.0%, greater than 0% and at most about 1.5%, greater than 0% and at most about 1.0%, greater than 0% and at most about 0.9%, greater than 0% and at most about 0.8%, greater than 0% and at most about 0.7%, 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 0.3%, greater than 0% and at most about 0.2%, or greater than 0% and at most about 0.1%.

[0028] The term "about" is used herein to provide literal support for the exact number it modifies, as well as numbers that are close to or approximate to the number modified by the term. In determining whether a number is close to or approximates a specifically stated number, a number that is close to or approximates a non-status-status number may be a number substantially equivalent to the number in the context in which it is presented. If the degree of approximation cannot be clearly determined by the context, "about" means within a range of plus or minus 10% of the provided value, or rounded to the nearest significant figure, in any case including the provided value. In some embodiments, the term "about" means a specified value ± at most 10%, at most ± 5%, or at most ± 1%.

[0029] In this method, if the percentage of cells expressing GREM1 in an enriched heterogeneous renal cell population is determined, the enriched heterogeneous renal cell population can be identified as having therapeutic potential when GREM1 is expressed in more than 0%, more than 0% and up to about 12%, more than 0% and up to about 10%, more than 0% and up to about 8%, more than 0% and up to about 7%, more than 0% and up to about 6%, more than 0% and up to about 5%, more than 0% and up to about 4%, more than 0% and up to about 3%, or more than 0% and up to about 2.5% of the cells in the enriched heterogeneous renal cell population.

[0030] In this method, if the percentage of cells expressing GREM1 in an enriched heterogeneous renal cell population is determined, the enriched heterogeneous renal cell population can be identified as having therapeutic potential when more than 5% and at most about 50%, more than 5% and at most about 45%, more than 5% and at most about 40%, more than 5% and at most about 35%, more than 5% and at most about 30%, more than 5% and at most about 25%, more than 5% and at most about 20%, more than 5% and at most about 15%, or more than 5% and at most about 10% of the cells in the enriched heterogeneous renal cell population express GREM1.

[0031] In this method, if the percentage of cells expressing ETV5 in an enriched heterogeneous renal cell population is determined, the enriched heterogeneous renal cell population can be identified as having therapeutic potential when more than 2%, more than about 2% and up to about 30%, more than about 2% and up to about 25%, more than about 2% and up to about 20%, more than 3%, more than about 3% and up to about 30%, more than about 3% and up to about 25%, more than about 3% and up to about 20%, more than 4%, more than about 4% and up to about 30%, more than about 4% and up to about 25%, and more than about 4% and up to about 20% of the cells in the enriched heterogeneous renal cell population express ETV5.

[0032] In this method, if the percentage of cells expressing ETV5 in an enriched heterogeneous renal cell population is determined, the enriched heterogeneous renal cell population can be identified as having therapeutic potential when it is determined that more than about 50% and at most about 60%, more than about 50% and at most about 65%, more than about 50% and at most about 70%, more than about 50% and at most about 75%, more than about 55% and at most about 60%, more than about 55% and at most about 65%, more than about 55% and at most about 70%, more than about 55% and at most about 75%, more than about 60% and at most about 65%, more than about 60% and at most about 70%, or more than about 60% and at most about 75% of the cells in the enriched heterogeneous renal cell population express ETV5.

[0033] In this method, if the percentage of HOX11-expressing cells in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than 1%, greater than about 1% and at most about 25%, greater than about 1% and at most about 20%, greater than about 1% and at most about 15%, greater than about 1% and at most about 12.5%, greater than about 2%, greater than about 2% and at most about 25%, greater than about 2% and at most about 20%, greater than about 2% and at most about 15%, greater than about 2% and at most about 12%, the percentage of cells expressing HOX11 in the enriched heterogeneous renal cell population is determined to be greater than 1%, greater than about 1 ...2%, the percentage of cells expressing HOX11 in the enriched heterogeneous renal cell population is determined to be greater than 1%, greater than about 1% and at most about 25%, greater than about 2% and at most about 20%, greater than about 2% and at most about 12%, the percentage of cells expressing HOX11 in the enriched heterogeneous renal cell population is determined to be greater than 1%, greater than about 1% and at most about 25%, greater than about 2% and at most about 12%, the percentage of cells expressing HOX11 in the enriched heterogeneous renal cell population is determined to be greater than 1%, greater than about 1% and at most about 25%, greater than about 2% and at most about 12 Cells expressing HOX11 at concentrations of 0.5%, greater than 2.5%, greater than about 2.5% and up to about 25%, greater than about 2.5% and up to about 20%, greater than about 2.5% and up to about 15%, greater than about 2.5% and up to about 12.5%, greater than about 3%, greater than about 3% and up to about 25%, greater than about 3% and up to about 20%, greater than about 3% and up to about 15%, or greater than about 3% and up to about 12.5% ​​can be identified as having therapeutic potential.

[0034] In this method, if the percentage of HOX11-expressing cells in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than 10%, greater than about 10% and at most about 20%, greater than about 10% and at most about 25%, greater than about 10% and at most about 30%, greater than about 10% and at most about 35%, or greater than about 15%, then the percentage is considered to be 15%. When up to about 20%, greater than about 15% and up to about 25%, greater than about 15% and up to about 30%, greater than about 15% and up to about 35%, greater than about 17.5% and up to about 20%, greater than about 17.5% and up to about 25%, greater than about 17.5% and up to about 30%, greater than about 17.5% and up to about 35%, greater than about 20% and up to about 25%, greater than about 20% and up to about 30%, or greater than about 20% and up to about 35% of cells express HOX11, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0035] In this method, if the percentage of NCAM1-expressing cells in an enriched heterogeneous renal cell population is determined, then the percentages of enriched heterogeneous renal cell populations are determined to be greater than 0%, greater than 0% and at most about 5%, greater than 0% and at most about 4%, 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 1.9%, greater than 0% and at most about 1.8%, greater than 0% and at most about 1.7%, greater than 0% and at most about 1.6%, greater than 0% and at most about 1.5%, and greater than about 0.25%. Cells expressing NCAM1 at concentrations greater than approximately 0.25% and up to approximately 5%, greater than approximately 0.25% and up to approximately 4%, greater than approximately 0.25% and up to approximately 3%, greater than approximately 0.25% and up to approximately 2.5%, greater than approximately 0.25% and up to approximately 2%, greater than approximately 0.25% and up to approximately 1.9%, greater than approximately 0.25% and up to approximately 1.8%, greater than approximately 0.25% and up to approximately 1.7%, greater than approximately 0.25% and up to approximately 1.6%, and greater than approximately 0.25% and up to approximately 1.5% can be identified as having therapeutic potential.

[0036] In this method, if the percentage of NCAM1-expressing cells in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than 0.5% and at most about 4%, greater than 0.5% and at most about 5%, greater than 0.5% and at most about 6%, greater than 0.5% and at most about 7%, greater than 0.5% and at most about 8%, greater than 0.5% and at most about 9%, greater than 0.5% and at most about 10%, greater than about 1% and at most about 4%, greater than about 1% and at most about 5%, greater than about 1% and at most about 6%, greater than about 1%, and greater than about 1%, the percentage of cells expressing NCAM1 is determined to be greater than 0.5% and at most about 4%, greater than about 1% and at most about 5%, greater than about 1% and at most about 6%, and greater than about 1%, the percentage of cells expressing NCAM1 is determined to be greater than 0.5% and at most about 4%, greater than about 0.5% and at most about 5%, greater than about 0.5 ... and greater than about 0.5% and at most about 6%, the percentage of cells expressing NCAM1 is determined to be greater than 0.5% and at most about 4%, greater than about 0.5% and at most about 0.5%, and greater than about 0.5% and at most about 0.5%, and greater than about 0.5% and at most Cells expressing NCAM1 at a rate of % and up to about 7%, greater than about 1% and up to about 8%, greater than about 1% and up to about 9%, greater than about 1% and up to about 10%, greater than about 1.5% and up to about 4%, greater than about 1.5% and up to about 5%, greater than about 1.5% and up to about 6%, greater than about 1.5% and up to about 7%, greater than about 1.5% and up to about 8%, greater than about 1.5% and up to about 9%, or greater than about 1.5% and up to about 10% can be identified as having therapeutic potential.

[0037] In this method, if the percentage of cells expressing THSD7A in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than about 0.5%, greater than about 0.5% and at most about 15%, greater than about 0.5% and at most about 12.5%, greater than about 0.5% and at most about 10%, greater than about 0.5% and at most about 8%, greater than about 0.5% and at most about 7%, greater than about 0.5% and at most about 6%, greater than about 0.5% and at most about 5%, greater than about 0.5% and at most about 4%, greater than about 1%, greater than about 1% and at most about 15%, greater than about 1% and at most about 12.5%, greater than about 1% and at most about 10%, greater than about 1% and at most about 8%, greater than about 1% and at most about 7%, greater than about 1% and at most about 6%, greater than about 1% and at most about 5%, greater than about 1% and at most about 6%, greater than about 1% and at most about 5%, greater than about 1% and at most about 1%, greater than about 1% and at most about 10%, greater than about 1% and at most about 8%, greater than about 1% and at most about 7%, greater than about 1% and at most about 6%, greater than about 1% and at most about 5%, greater than about 1% and at most about 1 ... When cells express THSD7A at approximately 4%, greater than approximately 1.5%, greater than approximately 1.5% and up to approximately 15%, greater than approximately 1.5% and up to approximately 12.5%, greater than approximately 1.5% and up to approximately 10%, greater than approximately 1.5% and up to approximately 8%, greater than approximately 1.5% and up to approximately 7%, greater than approximately 1.5% and up to approximately 6%, greater than approximately 1.5% and up to approximately 5%, greater than approximately 1.5% and up to approximately 4%, greater than approximately 2%, greater than approximately 2% and up to approximately 15%, greater than approximately 2% and up to approximately 12.5%, greater than approximately 2% and up to approximately 10%, greater than approximately 2% and up to approximately 8%, greater than approximately 2% and up to approximately 7%, greater than approximately 2% and up to approximately 6%, greater than approximately 2% and up to approximately 5%, or greater than approximately 2% and up to approximately 4%, enriched heterogeneous renal cell populations can be identified as having therapeutic potential.

[0038] In this method, if the percentage of cells expressing THSD7A in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than about 5% and at most about 50%, greater than about 5% and at most about 45%, greater than about 5% and at most about 40%, greater than about 5% and at most about 35%, greater than about 5% and at most about 30%, greater than about 5% and at most about 25%, greater than about 5% and at most about 20%, greater than about 10% and at most about 50%, greater than about 10% and at most about 45 ... When approximately 40%, greater than approximately 10% and up to approximately 35%, greater than approximately 10% and up to approximately 30%, greater than approximately 10% and up to approximately 25%, greater than approximately 10% and up to approximately 20%, greater than approximately 15% and up to approximately 50%, greater than approximately 45% and up to approximately 40%, greater than approximately 15% and up to approximately 35%, greater than approximately 15% and up to approximately 30%, greater than approximately 15% and up to approximately 25%, or greater than approximately 15% and up to approximately 20% of cells express THSD7A, the enriched heterogeneous population of kidney cells can be identified as having therapeutic potential.

[0039] In this method, if the percentage of cells expressing PTPRO in an enriched heterogeneous renal cell population is determined, then the percentages of enriched heterogeneous renal cell populations are determined to be greater than 0%, greater than 0% and at most about 5%, greater than 0% and at most about 4%, 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 1.9%, greater than 0% and at most about 1.8%, greater than 0% and at most about 1.7%, and greater than 0% and at most about 1.6%. Greater than 0% and at most about 1.5%, greater than about 0.1%, greater than about 0.1% and at most about 5%, greater than about 0.1% and at most about 4%, greater than about 0.1% and at most about 3%, greater than about 0.1% and at most about 2.5%, greater than about 0.1% and at most about 2%, greater than about 0.1% and at most about 1.9%, greater than about 0.1% and at most about 1.8%, greater than about 0.1% and at most about 1.7%, greater than about 0.1% and at most about 1.6%, greater than about 0.1% and at most about 1.5%, greater than Approximately 0.2%, greater than approximately 0.2% and at most approximately 5%, greater than approximately 0.2% and at most approximately 4%, greater than approximately 0.2% and at most approximately 3%, greater than approximately 0.2% and at most approximately 2.5%, greater than approximately 0.2% and at most approximately 2%, greater than approximately 0.2% and at most approximately 1.9%, greater than approximately 0.2% and at most approximately 1.8%, greater than approximately 0.2% and at most approximately 1.7%, greater than approximately 0.2% and at most approximately 1.6%, greater than approximately 0.2% and at most approximately 1.5%, greater than approximately 0.3%, greater than approximately 0.3% and at most approximately When cells express PTPRO at 5%, greater than about 0.3% and up to about 4%, greater than about 0.3% and up to about 3%, greater than about 0.3% and up to about 2.5%, greater than about 0.3% and up to about 2%, greater than about 0.3% and up to about 1.9%, greater than about 0.3% and up to about 1.8%, greater than about 0.3% and up to about 1.7%, greater than about 0.3% and up to about 1.6%, or greater than about 0.3% and up to about 1.5%, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0040] In this method, if the percentage of cells expressing PTPRO in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than 2% and at most about 15%, greater than 2% and at most about 14%, greater than 2% and at most about 13%, greater than 2% and at most about 12%, greater than 2% and at most about 11%, greater than 2% and at most about 10%, greater than 2% and at most about 9%, greater than 2% and at most about 8%, greater than 2% and at most about 7%, greater than 2% and at most about 6%, greater than 2% and at most about 5%, greater than about 2.5% and at most about 15%, greater than about 2.5% and at most about 14%, greater than about 2.5% and at most about 13%, greater than about 2.5% and at most about 12%, greater than about 2.5% and at most about 11%, etc., the percentage of cells expressing PTPRO is determined to be greater than 2%. When PTPRO is expressed in approximately 2.5% and up to approximately 10%, greater than approximately 2.5% and up to approximately 9%, greater than approximately 2.5% and up to approximately 8%, greater than approximately 2.5% and up to approximately 7%, greater than approximately 2.5% and up to approximately 6%, greater than approximately 2.5% and up to approximately 5%, greater than approximately 3% and up to approximately 15%, greater than approximately 3% and up to approximately 14%, greater than approximately 3% and up to approximately 13%, greater than approximately 3% and up to approximately 12%, greater than approximately 3% and up to approximately 11%, greater than approximately 3% and up to approximately 10%, greater than approximately 3% and up to approximately 9%, greater than approximately 3% and up to approximately 8%, greater than approximately 3% and up to approximately 7%, greater than approximately 3% and up to approximately 6%, or greater than approximately 3% and up to approximately 5%, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0041] In this method, if the percentage of cells expressing MPP5 in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than about 25%, greater than about 25% and at most about 75%, greater than about 25% and at most about 70%, greater than about 25% and at most about 65%, greater than about 25% and at most about 60%, greater than about 30%, greater than about 30% and at most about 75%, greater than about 30% and at most about 70%, greater than about 30% and at most about 65%, greater than about 30% and at most about 60%, greater than about 35%, greater than about 35% and at most about 75%, greater than about 35%, greater than about 35% and at most about 75%, greater than about 35%, greater than about 35%, greater than about 35%, greater than about 35%, greater than about 7 ... When MPP5 is expressed in approximately 35% to up to approximately 70%, greater than approximately 35% to up to approximately 65%, greater than approximately 35% to up to approximately 60%, greater than approximately 40%, greater than approximately 40% to up to approximately 75%, greater than approximately 40% to up to approximately 70%, greater than approximately 40% to up to approximately 65%, greater than approximately 40% to up to approximately 60%, greater than approximately 45%, greater than approximately 45% to up to approximately 75%, greater than approximately 45% to up to approximately 70%, greater than approximately 45% to up to approximately 65%, or greater than approximately 45% to up to approximately 60%, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0042] In this method, if the percentage of cells expressing MPP5 in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than about 70% and at most about 99%, greater than about 70% and at most about 95%, greater than about 70% and at most about 90%, greater than about 70% and at most about 85%, greater than about 70% and at most about 80%, greater than about 75% and at most about 99%, greater than about 75% and at most about 95%, greater than about 75% and at most about 90%, greater than about 75% and at most about 85%, the percentage of cells expressing MPP5 in an enriched heterogeneous renal cell population is determined to be greater than about 70% and at most about 99%, greater than about 75% and at most about 95%, greater than about 75% and at most about 85%, the percentage of cells expressing MPP5 in an enriched heterogeneous renal cell population is determined to be greater than about 70% and at most about 99%, greater than about 75% and at most about 95%, greater than about 75% and at most about 85%, the percentage of cells expressing MPP5 in an enriched heterogeneous renal cell population is determined to be greater than about 70% and at most about 99%, greater than about 75% and at most about 95%, greater than about 75% and at most about 85%, the percentage of cells expressing MPP5 in an enriched heterogeneous renal cell population is determined to be greater than about 70% and at most about 99%, greater than about 70 ... When cells express MPP5, enriched heterogeneous renal cell populations can be identified as having therapeutic potential.

[0043] In this method, if the percentage of cells expressing TPPP3 in an enriched heterogeneous renal cell population is determined, then when the enriched heterogeneous renal cell population is determined to be greater than 0%, greater than 0% and at most about 5%, greater than 0% and at most about 4%, greater than 0% and at most about 3%, greater than 0% and at most about 2%, greater than 0% and at most about 1%, greater than 0% and at most about 0.9%, greater than 0% and at most about 0.8%, greater than 0% and at most about 0.7%, 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 about 0.1%, greater than about 0.1% and at most about 5%, greater than about 0.1% and at most about 4%, greater than about 0.1% and at most about 3%, greater than about 0.1% and at most about 2%, greater than about 0.1% and at most about 1%, greater than about 0.1% and at most about 0.1%, the percentage of cells expressing TPPP3 in the enriched heterogeneous renal cell population is determined to be greater than 0%, greater than 0% and at most about 5%, greater than 0% and at most about 4%, greater than about 0.1% and at most about 3%, greater than about 0.1% and at most about 2%, greater than about 0.1% and at most about 1%, greater than about 0.1% and at most about 0%. When cells express TPPP3, enriched heterogeneous renal cell populations can be identified as having therapeutic potential.

[0044] In this method, if the percentage of cells expressing LMX1B in an enriched heterogeneous renal cell population is determined, the enriched heterogeneous renal cell population can be identified as having therapeutic potential when more than 0%, more than 0% and at most about 5%, more than 0% and at most about 4%, more than 0% and at most about 3%, more than 0% and at most about 2%, more than 0% and at most about 1%, more than 0% and at most about 0.9%, more than 0% and at most about 0.8%, more than 0% and at most about 0.7%, more than 0% and at most about 0.6%, more than 0% and at most about 0.5%, more than 0% and at most about 0.4%, more than 0% and at most about 0.3%, more than 0% and at most about 0.25%, or more than 0% and at most about 0.2% of the cells in the enriched heterogeneous renal cell population express LMX1B.

[0045] For example, in an example of a method for further determining the percentage of cells expressing at least one (any one, at least two, at least three, at least four, or five) UB / CM markers in an enriched heterogeneous renal cell population, an enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all of the following are determined: (i) more than 0% of the cells in the enriched heterogeneous renal cell population express WNT11; (ii) more than 0% of the cells in the enriched heterogeneous renal cell population express GREM1; (iii) more than about 2% and at most about 30% of the cells in the enriched heterogeneous renal cell population express ETV5; (iv) more than about 1% and at most about 25% of the cells in the enriched heterogeneous renal cell population express HOX11; and / or (v) more than 0% of the enriched heterogeneous renal cell population expresses NCAM1. In another example of a method for further determining the percentage of cells expressing at least one (any one, at least two, at least three, at least four, or five) UB / CM markers in an enriched heterogeneous renal cell population, an enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all of the following are determined: (i) greater than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11; (ii) greater than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1; (iii) greater than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5; (iv) greater than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11 and / or (v) greater than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1.

[0046] In an example of a method for further determining the percentage of cells expressing at least one (any one, at least two, at least three, at least four, or five) UB / CM markers in an enriched heterogeneous renal cell population, an enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all of the following are determined: (i) more than 0% of the cells in the enriched heterogeneous renal cell population express WNT11; (ii) more than about 5% and at most about 50% of the cells in the enriched heterogeneous renal cell population express GREM1; (iii) more than about 50% and at most about 75% of the cells in the enriched heterogeneous renal cell population express ETV5; (iv) more than about 10% and at most about 35% of the cells in the enriched heterogeneous renal cell population express HOX11; and / or (v) more than about 0.5% and at most about 10% of the enriched heterogeneous renal cell population express NCAM1. In another example of a method for further determining the percentage of cells expressing at least one (any one, at least two, at least three, at least four, or five) UB / CM markers in an enriched heterogeneous renal cell population, an enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all of the following are determined: (i) greater than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11; (ii) greater than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1; (iii) greater than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5; (iv) greater than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11 and / or (v) greater than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1.

[0047] Any combination of any two, three, four, or all five markers expressed, which can be further determined at these percentages, can be any of the following combinations: WNT11 and GREM1; WNT11 and ETV5; WNT11 and HOX11; WNT11 and NCAM1; GREM1 and ETV5; GREM1 and HOX11; GREM1 and NCAM1; ETV5 and HOX11; ETV5 and NCAM1; HOX11 and NCAM1; WNT11, GREM1 and ETV5; WNT11, GREM1 and HOX11; WNT11, GREM1 and NCAM1; WNT11, ETV5 and HOX11; WNT11 ETV5 and NCAM1; WNT11, HOX11 and NCAM1; GREM1, ETV5 and HOX11; GREM1, ETV5 and NCAM1; GREM1, HOX11 and NCAM1; ETV, HOX11 and NCAM1; WNT11, GREM1, ETV5 and HOX11; WNT11, GREM1, ETV5 and NCAM1; WNT11, ETV5, HOX11 and NCAM1; WNT11, GREM1, HOX11 and NCAM1; GREM1, ETV5, HOX11 and NCAM1; or WNT11, GREM1, ETV5, HOX11 and NCAM1.

[0048] In some instances of these methods, in addition to determining the percentage of cells expressing any two, three, four, or all five UB / CM markers in an enriched heterogeneous renal cell population to identify it as having therapeutic potential, it may also determine whether the cells in the enriched heterogeneous renal cell population express at least one (any one, at least two, at least three, at least four, or five) podocyte markers to identify it as having therapeutic potential. Alternatively, in addition to determining the percentage of cells expressing any two, three, four, or all five UB / CM markers in an enriched heterogeneous renal cell population to identify it as having therapeutic potential, it may also determine the percentage of cells expressing at least one (any one, at least two, at least three, at least four, or five) podocyte markers in the enriched heterogeneous renal cell population to identify it as having therapeutic potential.

[0049] For example, in an example of a method for further determining the percentage of cells expressing at least one (any one, at least two, at least three, at least four, or five) podocyte markers in an enriched heterogeneous renal cell population, an enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all of the following are determined: (i) more than about 0.5% and at most about 15% of the enriched heterogeneous renal cell population express THSD7A; (ii) more than 0% of the enriched heterogeneous renal cell population express PTPRO; (iii) more than about 25% and at most about 75% of the enriched heterogeneous renal cell population express MPP5; (iv) more than 0% of the enriched heterogeneous renal cell population express TPPP3 and / or (v) more than 0% of the enriched heterogeneous renal cell population express LMX1B. In another example of a method for further determining the percentage of cells expressing at least one (at least one, at least two, at least three, at least four, or five) podocyte markers in an enriched heterogeneous renal cell population, 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 about 1.5% and at most about 6% of the enriched heterogeneous renal cell population express THSD7A; (ii) greater than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population express PTPRO; (iii) greater than about 40% and at most about 70% of the enriched heterogeneous renal cell population express MPP5; (iv) greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population express TPPP3; and / or (v) greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population express LMX1B.

[0050] In an example of a method for further determining the percentage of cells expressing at least one (any one, at least two, at least three, at least four, or five) podocyte markers in an enriched heterogeneous renal cell population, an enriched heterogeneous renal cell population may be identified as having therapeutic potential if any two, any three, any four, or all of the following are determined: (i) more than about 5% and at most about 50% of the enriched heterogeneous renal cell population express THSD7A; (ii) more than 2% and at most about 15% of the enriched heterogeneous renal cell population express PTPRO; (iii) more than about 70% and at most about 99% of the enriched heterogeneous renal cell population express MPP5; (iv) more than 0% of the enriched heterogeneous renal cell population express TPPP3 and / or (v) more than 0% of the enriched heterogeneous renal cell population express LMX1B. In another example of a method for further determining the percentage of cells expressing at least one (at least one, at least two, at least three, at least four, or five) podocyte markers in an enriched heterogeneous renal cell population, 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 about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A; (ii) greater than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO; (iii) greater than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5; (iv) greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3; and / or (v) greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B.

[0051] Any combination of any two, three, four, or all five markers expressed, which can be further determined at these percentages, can be any of the following combinations: THSD7A and PTPRO; THSD7A and MPP5; THSD7A and TPPP3; THSD7A and LMX1B; PTPRO and MPP5; PTPRO and TPPP3; PTPRO and LMX1B; MPP5 and TPPP3; MPP5 and LMX1B; TPPP3 and LMX1B; THSD7A, PTPRO, and MPP5; THSD7A, PTPRO, and TPPP3; THSD7A, PTPRO, and LMX1B; THSD7A, MPP5, and TPPP3; THSD7 A, MPP5 and LMX1B; THSD7A, TPPP3 and LMX1B; PTPRO, MPP5 and TPPP3; PTPRO, MPP5 and LMX1B; PTPRO, TPPP3 and LMX1B; MPP5, TPPP3 and LMX1B; THSD7A, PTPRO, MPP5 and TPPP3; THSD7A, PTPRO, MPP5 and LMX1B; THSD7A, MPP5, TPPP3 and LMX1B; THSD7A, PTPRO, TPPP3 and LMX1B; GREM1, MPP5, TPPP3 and LMX1B; or THSD7A, PTPRO, MPP5, TPPP3 and LMX1B.

[0052] In some instances of these methods, in addition to determining the percentage of cells expressing any two, three, four, or all five podocyte markers in an enriched heterogeneous renal cell population to identify the enriched heterogeneous renal cell population as having therapeutic potential, it may also determine whether the cells in the enriched heterogeneous renal cell population express at least one (any one, at least two, at least three, at least four, or five) UB / CM marker to identify the enriched heterogeneous renal cell population as having therapeutic potential.

[0053] In methods for identifying enriched heterogeneous renal cell populations as having therapeutic potential, in addition to determining whether the cells in the enriched heterogeneous renal cell population express (i) at least one (any one, at least two, at least three, at least four or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte markers, the expression of at least one additional marker may also be detected. At least one additional biomarker may be another UB / CM biomarker, such as rearranged during transfection (RET), fibroblast growth factor (FGF) 8, FGF 10, cbp / P300 interacting transactivator (CITED) 1, eyes absent (EYA) 1, sineoculis homeobox homolog 2 (SIX2), odd-skipped related 1 (OSR1), and / or glial cell derived neurotrophic factor (GDNF). Determining the expression of at least one additional biomarker and (i) at least one (any one, at least two, at least three, at least four or five) UB / CM biomarkers; and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte biomarkers can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0054] In a method 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 (i) at least one (any one, at least two, at least three, at least four, or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four, or five) podocyte markers, and optionally, (iii) one or more other markers, it may additionally determine whether the cells of the enriched heterogeneous renal cell population express at least one additional marker, wherein the at least one additional marker may be an additional podocyte marker, such as nphs1 (podin). Determining the expression of at least one additional marker and (i) at least one (any one, at least two, at least three, at least four, or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four, or five) podocyte markers, and optionally, (iii) one or more other markers, may identify the enriched heterogeneous renal cell population as having therapeutic potential.

[0055] 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 (i) at least one (any one, at least two, at least three, at least four, or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four, or five) podocyte markers; and optionally, (iii) one or more additional and / or supplementary markers, it may additionally determine whether the cells of the enriched heterogeneous renal cell population express at least one podocyte-PEC marker. The podocyte-PEC marker may be or include hes1 or CD44. Determining the expression of at least one podocyte-PEC marker and (i) at least one (any one, at least two, at least three, at least four or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte markers, and optionally, (iii) at least one or more additional and / or supplementary markers, can identify enriched heterogeneous renal cell populations as having therapeutic potential.

[0056] In an example of a method for further determining whether cells in an enriched heterogeneous renal cell population express the podocyte-PEC marker, and where the podocyte-PEC marker is hes1, determining whether cells in an enriched heterogeneous renal cell population express hes1 could be determining a certain percentage of cells in the enriched heterogeneous renal cell population that express hes1. If, in an example of this method, a certain percentage of cells in the enriched heterogeneous renal cell population are further determined to express hes1, then the enriched heterogeneous renal cell population can be identified as having therapeutic potential when it is further determined that greater than about 65%, greater than about 65% and up to about 98%, greater than about 65% and up to about 95%, greater than about 65% and up to about 90%, greater than about 70%, greater than about 70% and up to about 98%, greater than about 70% and up to about 95%, greater than about 70% and up to about 90%, greater than about 75%, greater than about 75% and up to about 98%, greater than about 75% and up to about 95%, greater than about 75% and up to about 90%, greater than about 80%, greater than about 80% and up to about 98%, greater than about 80% and up to about 95%, or greater than about 80% and up to about 90% of the cells in the enriched heterogeneous renal cell population express hes1.

[0057] In addition, in the method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, besides determining whether the cells of the enriched heterogeneous renal cell population express (i) at least one (any one, at least two, at least three, at least four or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte markers and, optionally, (iii) one or more additional and / or supplementary markers, and optionally, (iv) at least one podocyte-PEC marker, it may also determine whether the cells of the enriched heterogeneous renal cell population express transforming growth factor β2 (TGFβ2). If, in addition to at least one (any one, at least two, at least three, at least four or five) UB / CM markers; and / or (ii) at least one (any one, at least two, at least three, at least four or five) podocyte markers, and optionally, (iii) at least one or more additional and / or supplementary markers, and optionally, (iv) at least one podocyte-PEC marker, the enriched heterogeneous renal cell population also expresses TGFβ2, then the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0058] In an example of a method for further determining whether cells in an enriched heterogeneous renal cell population express TGFβ2, determining whether cells in the enriched heterogeneous renal cell population express TGFβ2 can be done by measuring a certain percentage of cells in the enriched heterogeneous renal cell population that express TGFβ2, or by measuring whether cells in the enriched heterogeneous renal cell population secrete a certain amount of TGFβ2. In these cases, if a certain percentage of cells express TGFβ2 or a certain amount of TGFβ2 is further measured, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0059] If, in an example of this method, a certain percentage of cells in the enriched heterogeneous renal cell population are further determined to express TGFβ2, then when it is determined that the enriched heterogeneous renal cell population is greater than 40%, 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%, greater than 45%, 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 When approximately 45% to at most approximately 70%, greater than 50%, greater than approximately 50% to at most approximately 85%, greater than approximately 50% to at most approximately 80%, greater than approximately 50% to at most approximately 75%, greater than approximately 50% to at most approximately 70%, greater than 55%, greater than approximately 55% to at most approximately 85%, greater than approximately 55% to at most approximately 80%, greater than approximately 55% to at most approximately 75%, or greater than approximately 55% to at most approximately 80% of cells express TGFβ2, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0060] If, in an example of this method, the amount of TGFβ2 secreted by the enriched heterogeneous renal cell population is further determined, then when it is determined that the enriched heterogeneous renal cell population secretes more than about 1 ng, or more than about 1.2 ng, or more than about 1.4 ng, or more than about 1.6 ng, or more than about 1.8 ng, or more than about 2 ng, or more than about 2.2 ng, or more than about 2.4 ng, or more than about 2.6 ng, or more than about 2.8 ng, or more than about 3.0 ng, or more than about 3.5 ng, or more than about 4.0 ng, or more than about 4.5 ng, or more than about 5.0 ng, or more than about 5.5 ng, or more than about 6.0, or more than about 6.5 ng, or more than about 7.0 ng, or more than about 7.5 ng, or more than about 8.0 ng, or more than about 8.5 ng per million cells, the amount of TGFβ2 secreted by the cells is determined to be greater than about 1 ng, or more than about 1.2 ng, or more than about 1.4 ng, or more than about 1.6 ng, or more than about 1.8 ng, or more than about 2.0 ng, or more than about 2.0 ng, or more than about 4.0 ng, or more than about 4.5 ng, or more than about 5.0 ng, or more than about 5.0 ng, or more than about 5.5 ng, or more than about 6.0, or more than about 6.5 ng, or more than about 7.0 ng, or more than about 7.5 ng, or more than about 8.0 ng, or more than about 8.5 ng. Enriched heterogeneous renal cell populations can be identified as having therapeutic potential when TGFβ2 levels are greater than or equal to about 9.0 or about 9.5 ng. TGFβ2 secreted by these enriched heterogeneous renal cell populations in these amounts can be in culture medium within time periods 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, about 24 hours to about 48 hours, or about 48 hours to about 72 hours.

[0061] In another method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, as provided in this disclosure, the percentage of renal cells expressing UB, CM, and podocyte characteristic genes in the enriched heterogeneous renal cell population can be determined. An enriched heterogeneous renal cell population can be identified as having therapeutic potential if: (i) approximately 20% to approximately 40% of the renal cells in the population express ureteral bud and cap mesenchymal cell characteristic genes; and (ii) approximately 50% to approximately 65% ​​of the renal cells in the population express podocyte characteristic genes. In these methods, the renal cell populations are prepared from non-fetal renal tissue.

[0062] In such methods of this disclosure, if (i) in the group approximately 15% to approximately 50%, approximately 15% to approximately 48%, approximately 15% to approximately 46%, approximately 15% to approximately 44%, approximately 15% to approximately 42%, approximately 15% to approximately 40%, approximately 17.5% to approximately 50%, approximately 17.5% to approximately 48%, approximately 17.5% to approximately 46%, approximately 17.5% to approximately 44%, approximately 17.5% to approximately 42%, approximately 17.5% to approximately 40%, approximately 20% to approximately 50%, approximately 20% to approximately 48%, approximately 20% to approximately 46%, approximately 20% to approximately 44%, approximately 20% to approximately 42% or about 20% to about 40% of the cells express ureteral bud and cap mesenchymal cell characteristic genes; and (ii) about 45% to about 75%, about 45% to about 70%, about 45% to about 65%, about 45% to about 60%, about 50% to about 75%, about 50% to about 70%, about 50% to about 65%, about 50% to about 60%, about 55% to about 75%, about 55% to about 70%, about 55% to about 65%, or about 55% to about 60% of the cells in the population express podocyte characteristic genes, thus identifying heterogeneous renal cell populations as having therapeutic potential.

[0063] In some instances of these methods, a heterogeneous renal cell population can be identified as having therapeutic potential if (i) approximately 15% to approximately 50% of the cells in the population express ureteric bud and cap-mesenchymal cell characteristic genes and (ii) approximately 45% to approximately 75% of the cells in the population express podocyte characteristic genes. In other instances of these methods, a heterogeneous renal cell population can be identified as having therapeutic potential if (i) approximately 17.5% to approximately 46% of the cells in the population express ureteric bud and cap-mesenchymal cell characteristic genes and (ii) approximately 45% to approximately 70% of the cells in the population express podocyte characteristic genes. In still other instances, a heterogeneous renal cell population can be identified as having therapeutic potential if (i) approximately 20% to approximately 40% of the renal cells in the population express ureteric bud and cap-mesenchymal cell characteristic genes and (ii) approximately 50% to approximately 65% ​​of the renal cells in the population express podocyte characteristic genes. In other instances, heterogeneous renal cell populations can be identified as having therapeutic potential if (i) approximately 20% to approximately 40% of the renal cells in a population express ureteral bud and cap mesenchymal cell characteristic genes and (ii) approximately 55% to approximately 65% ​​of the renal cells in a population express podocyte characteristic genes.

[0064] In a method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, the percentage of renal cells expressing UB, CM, and podocyte-specific genes in the enriched heterogeneous renal cell population is determined. UB and CM cell-specific genes may include or include Wingless-associated integration site family member 11 (WNT11), Gremlin (GREM)1, E26 transformation-specific (ETS) variant transcription 5 (ETV5), homeobox (HOX)11, neural cell adhesion molecule (NCAM)1, transfection-rearranged (RET), fibroblast growth factor 8 (FGF8), FGF10, cbp / P300 interacting transactivator (CITED)1, eye-agnostic (EYA)1, sine oculis homeobox homologue 2 (SIX2), Odd-skipped associated protein 1 (OSR1), and glial cell-derived neurotrophic factor (GDNF). Alternatively, the characteristic genes of UB and CM cells consist of WNT11, GREM1, ETV5, HOX11, NCAM1, RET, FGF8, FGF10, CITED1, EYA1, SIX2, OSR1, and GDNF.

[0065] In a method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, the percentage of renal cells expressing UB, CM, and podocyte-specific genes within the enriched heterogeneous renal cell population can be determined. The podocyte-specific genes may be or include matrix metalloproteinase (MPP) 5. Alternatively, the podocyte-specific genes may be or include MPP5 and one or more of the following: platelet-reactive protein type 1 domain protein (THSD) 7A, protein tyrosine phosphatase receptor type O (PTPRO), tubulin polymerization-promoting protein (TPPP) 3, lim homeobox transcription factor 1β (LMX1B), and / or nephrin (NPHS1). Alternatively, the podocyte-specific genes may include THSD7A, PTPRO, MPP5, TPPP3, LMX1B, and NPHS1, or may consist of THSD7A, PTPRO, MPP5, TPPP3, LMX1B, and NPHS1.

[0066] In a method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, the percentage of renal cells expressing UB, CM, and podocyte-specific genes within the enriched heterogeneous renal cell population can be determined. The enriched heterogeneous renal cell population can have been prepared from non-fetal renal tissue. The non-fetal renal tissue can be from the kidney of a subject or from a renal tissue block, such as a biopsy. The subject can be of any age, for example, older than 1 month of age.

[0067] It should be understood that if the percentage of cells expressing a certain biomarker or a gene characteristic of UB / CM / podocytes is provided as a percentage “about” a specific number, such as about 5%, then the percentage of cells does not need to be precisely that specific number, for example, exactly 5%. Rather, it should be understood that the percentage of cells can be within up to 10% of that specific number, for example, between 4.5% and 5.5%. It should also be understood that if cells express a biomarker by secreting it in an amount “about” a specific number (e.g., about 5 ng), then that amount does not need to be precisely that specific number, for example, exactly 5 ng. Rather, it should be understood that the amount of biomarker secreted can be within 10% of that specific number, for example, between 4.5 and 5.5 ng.

[0068] In any method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, determining whether the cells in the enriched heterogeneous renal cell population express any markers, such as UB / CM markers (WNT11, GREM1, ETV5, HOX11, NCAM1, RET, FGF8, FGF10, CITED1, EYA1, SIX2, OSR1, and / or GDNF), podocyte markers (THSD7A, PTPRI, MMP5, TPPP3, LIMX1B, and / or NPHS1), podocyte-PEC markers (hes1 or CD44), or added markers (TGFβ2), can be performed by detecting markers in nucleic acid (e.g., mRNA or miRNA), peptide form, or both nucleic acid and peptide forms, and can be detected by any suitable assay / technique. For example, if expression is determined in peptide form, it can be detected by assays such as Western blotting, fluorescence-activated cell sorting (FACS), enzyme-linked immunosorbent assay (ELISA). Immunosorbent assay (ELISA) is used for determination. 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 serialization analysis (SAGE), mass spectrometry array (MassARRAY), or fluorescence in situ hybridization (FISH), or by using Southern blotting, polymerase chain reaction (PCR) or reverse transcriptase PCR, gene expression serialization analysis (SAGE), mass spectrometry array, or fluorescence in situ hybridization (FISH). Expression can be determined by bulk or single-cell assays, or after collecting conditioned medium from cell samples enriched with heterogeneous kidney cell populations. Assays can be assays using labeled detection reagents. 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 131 I. Colloidal gold particles, fluorescent markers such as Texas Red, Rhodamine, fluorescein, dansyl chloride, lissamine, phycoerythrin, phycocyanin, SPECTRUM ORANGE, SPECTRUM GREEN1, and enzyme substrates such as firefly luciferase, bacterial luciferase, luciferin, horseradish peroxidase, alkaline phosphatase, or β-galactosidase.

[0069] The enriched heterogeneous renal cell population 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, and the enriched heterogeneous renal cell population may be identified as having therapeutic potential by any method. This enrichment of one or more renal cell types in the enriched heterogeneous renal cell population can refer to a higher percentage of one or more renal cell types compared to a patient's renal tissue, a patient's renal biopsy, or an in vitro culture of cells established from a patient's renal tissue or renal biopsy (these may be collectively referred to as the "initial renal cell population"). In the case of an in vitro culture of cells established from a patient's renal tissue or a patient's renal biopsy, the initial renal cell population may be a renal cell preparation containing renal tissue or renal biopsy dissociated cells (e.g., cells dissociated from renal tissue or renal biopsy via mincing 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 enriched 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. Within this enriched heterogeneous renal cell population, these cell types may be able to express and can express markers expressed by UB, CM, and podocytes.

[0070] One or more kidney cell types can be enriched from an enriched heterogeneous kidney cell population via a method that includes a separation step, resulting from preparation of the initial kidney cell population (e.g., a patient's kidney tissue, a patient's kidney biopsy, or an in vitro culture of cells established from the patient's kidney tissue or kidney biopsy). The separation step can be a buoyancy-density-based separation step of the initial kidney cell population, which has been passaged no more than once, twice, or three times. If the separation step is a buoyancy-density-based separation step, the separation step can 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 at most once, twice, or three times) into one or more distinguishable fractions from which the enriched heterogeneous kidney cell population can be clearly identified and isolated. A distinguishable fraction may be a fraction 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 may be a fraction 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 may be a fraction in which the buoyant density is between about 1.045 g / mL and about 1.091 g / mL, or between about 1.045 g / mL and about 1.052 g / mL. Alternatively, the separation step can be a separation step that separates cells from the starting kidney cell population (or cells from the starting kidney cell population that have been passaged no more than once, twice, or three times) based on whether the cells express specific markers on their surface. If the separation step separates cells based on the expression of specific cell surface markers, then the separation step can be a separation step using flow cytometry. If the cells express characteristic specific surface markers, such as those characteristic of renal epithelium, renal tubules, renal tubular epithelium, or proximal and / or distal tubular cells, such as cytokeratins, such as CK8, CK18, and / or CK19, then flow cytometry can sort these cells from the starting kidney cell population.

[0071] A heterogeneous population of kidney cells enriched prior to the separation step can be cultured under hypoxic conditions. This population has been prepared from a starting kidney cell population (or a starting kidney cell population that has been passaged one, two, or three times). If cells are cultured under hypoxic conditions prior to the separation step, they can be cultured under conditions where the oxygen level is 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% oxygen. 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 to about 48 hours, about 6 to about 36 hours, about 6 to about 24 hours, about 12 to about 48 hours, about 12 to about 36 hours, about 12 to about 24 hours, or about 12 to about 18 hours.

[0072] Typically, enriched heterogeneous renal 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 a patient's renal biopsy. If the enriched heterogeneous renal cell population is prepared from an in vitro culture of cells established from a patient's kidney tissue or a 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 a 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 renal cell population can be isolated by performing a separation step. Once the enriched heterogeneous renal cell population has been isolated, it can be frozen and / or analyzed before being used as a therapeutic agent.

[0073] If an enriched heterogeneous population of kidney cells 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 kidney disease in a patient with such need, and / or used to prepare a medicament for treating kidney disease. If the enriched heterogeneous population of kidney cells is identified as having therapeutic potential and included in a pharmaceutical composition, it may be formulated as a hydrogel composition or a liquid composition. The pharmaceutical composition may or may not contain hyaluronic acid.

[0074] If the pharmaceutical composition is formulated into a hydrogel composition, the cells of the enriched heterogeneous kidney cell composition can be combined with temperature-sensitive cell-stabilizing biomaterials. These temperature-sensitive cell-stabilizing biomaterials can be biomaterials that are 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-to-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-to-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-to-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 one or more naturally derived or recombinant proteins or peptides, 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 αI collagen, such as porcine type I αI collagen or recombinant human type I αI collagen. 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 renal cell population may be dispersed throughout the biomaterial or substantially uniformly distributed throughout the biomaterial. The enriched heterogeneous renal 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 10-1. 6 cells / mL, approximately 40 x 10⁻⁶ 6 cells / mL, approximately 60 x 10⁻⁶ 6 cells / mL, approximately 100 x 10⁻⁶ 6 cells / mL, approximately 120 x 10⁻⁶ 6 cells / mL, approximately 140 x 10⁻⁶ 6 cells / mL, approximately 160 x 10⁻⁶ 6 cells / mL, approximately 180 x 10⁻⁶ 6 1 cell / mL or approximately 200 x 10⁻⁶ 6 Cells / mL.

[0075] If the pharmaceutical composition is formulated as a liquid composition, the enriched heterogeneous renal 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 renal 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 renal 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 102 6 cells / mL, approximately 40 x 10⁻⁶ 6 cells / mL, approximately 60 x 10⁻⁶ 6 cells / mL, approximately 100 x 10⁻⁶ 6 cells / mL, approximately 120 x 10⁻⁶ 6 cells / mL, approximately 140 x 10⁻⁶ 6 cells / mL, approximately 160 x 10⁻⁶ 6 cells / mL, approximately 180 x 10⁻⁶ 6 1 cell / mL or approximately 200 x 10⁻⁶ 6 Cells / mL.

[0076] If an enriched heterogeneous renal cell population is identified as having therapeutic potential, the enriched heterogeneous renal cell population or a pharmaceutical composition containing such a 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 containing such a population may be used to prepare a medicament for treating kidney disease. Kidney disease can be at any stage or extent of acute or chronic renal failure. 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, for example, erythropoietin deficiency, and mineral imbalances, such as vitamin D deficiency.

[0077] If an enriched heterogeneous population of renal cells is identified as having therapeutic potential, the administration of the enriched heterogeneous renal cell population or a pharmaceutical composition containing such a population can treat kidney disease. It can treat kidney disease by restoring renal function, stabilizing renal function, improving renal function, reducing renal fibrosis, or reducing renal inflammation in the kidneys of patients requiring such treatment. Treatment of kidney disease can restore mineral balance, electrolyte balance, fluid homeostasis, reabsorption of essential nutrients, or cystatin C metabolism, or alleviate anemia in patients requiring such treatment. Treatment of kidney disease can delay or avoid the need for dialysis, or it can delay or avoid the need for kidney transplantation in patients requiring treatment for kidney disease. If treatment of kidney disease delays a patient's need for dialysis or kidney transplantation, this 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 determined 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, and glomerular filtration rate (GFR), weight, blood pressure (mean general blood pressure, diastolic blood pressure, or systolic blood pressure), and physical endurance performance.

[0078] 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 containing 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 containing an enriched heterogeneous renal cell population can be administered to or into the kidney of a patient requiring treatment for kidney disease. If the enriched heterogeneous renal cell population is administered to or into the kidney of a patient requiring treatment for kidney disease, it can be administered via single or multiple injections. It can be administered via direct laparotomy, via direct laparoscopy, transabdominal, or percutaneous. The enriched heterogeneous renal cell population or a pharmaceutical composition containing an enriched heterogeneous renal cell population can be administered percutaneously into the renal cortex, or it can be administered by percutaneously inserting a guiding cannula to puncture the renal capsule and then injecting the enriched heterogeneous renal cell population into the kidney. Enriched heterogeneous renal cell populations or pharmaceutical compositions containing enriched heterogeneous renal cell populations can be administered by injection into the renal cortex of one or both kidneys of a patient. Administration of enriched heterogeneous renal cell populations or pharmaceutical compositions containing enriched heterogeneous renal cell populations 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.

[0079] Enriched heterogeneous renal cell populations or pharmaceutical compositions containing enriched heterogeneous renal cell populations may be administered at a therapeutically effective dose via any suitable route. A therapeutically effective dose or amount administered to a patient requiring treatment for kidney disease may include approximately 1-9 x 10^6 cells per gram of the patient's estimated kidney weight. 6 A heterogeneous population of enriched renal cells. The therapeutically effective dose of the drug composition may be approximately 1.0 x 10⁻⁶ cells per gram of the patient's estimated kidney weight. 6 Approximately 2.0 x 10 6 Approximately 3.0 x 10 6、约 4.0x10 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 –7.0x10 6 A dose of enriched heterogeneous renal cell populations.

[0080] In methods of treating kidney disease, administration of a therapeutic composition comprising a heterogeneous population of renal cells to a patient can be performed via a first and second injection. The first and second injections can be administered approximately 3 to 12 months apart. Specifically, the intervals between the first and second injections can be approximately 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months. Alternatively, the intervals between the first and second injections can be approximately 3 to 6 months, 6 to 9 months, 9 to 12 months, 3 to 9 months, 6 to 9 months, or 6 to 12 months.

[0081] In some implementations, the enriched heterogeneous kidney cell population can be cultured in a three-dimensional (D) system until it forms organoids, such as cell clusters, which may have been identified as having therapeutic potential or may not yet have been identified as having therapeutic potential. The 3D culture system can be a suspension culture. In the case of a suspension culture system, it may include a stirrer and lack a scaffold for the attachment of the enriched heterogeneous kidney cell population. The cells of the enriched heterogeneous kidney cell population can be cultured in the 3D culture system for approximately 7 to approximately 18 days to form organoids. The cells of the enriched heterogeneous kidney cell population can be cultured in the 3D culture system for approximately 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 days to form organoids. The organoids formed in the 3D culture system can have approximately 100... m to approximately 800 The diameter is m. Organoids formed in 3D culture systems can have a diameter of approximately 100 m. m to approximately 600 m, or about 100 m to approximately 400, or approximately 100 m to approximately 200 m, or about 200 m to approximately 800 m, or about 400 m to approximately 800 m, or about 600 m to approximately 800 The diameter of m.

[0082] Organoids prepared from enriched heterogeneous kidney cell populations in a 3D culture system can possess certain characteristics. One of these characteristics may be the expression of certain biomarkers, such as nucleic acid and protein biomarkers. Certain biomarkers that can be expressed from the cells of organoids may include any one or more (any one, any two, any three, any four, any five, any six, any seven, or all eight) of Hoxa11, CK8, Meis2, Thsd7a, Ptpro, CD13, LTL, and / or Cdc6. In some cases, the markers may be (i) Hoxa11 and Meis2, or (ii) Hoxa11, Meis2, and CK8, or (iii) Thsd7a and Ptpro, or (iv) CD13 and LTL, or (v) Cdc6, or (vi) Hoxa11, Meis2, Thsd7a, and Ptpro, or (vii) Cdc6, Hoxa11, Meis2, and Thsd7A, or (viii) Hoxa11 and Thsd7A, or (ix) Cdc6, Hoxa11, and Thsd7A, or (x) Ptpro, or (xi) Meis2, or (xii) Thsd7A. Organoids prepared from enriched heterogeneous renal cell populations may also, or alternatively, exhibit certain structural features. Some of the structural features exhibited may include nephron-like structures, for example, structures with tubular and / or glomerular features. In some cases, organoid cells can form structures exhibiting a glomerular-like appearance. Structures exhibiting a glomerular-like appearance may include structures with PEC-like margins. Such structural features can be exhibited by organoids at or after approximately 6 to approximately 10 days of culture (e.g., approximately 6, 7, 8, 9, or 10 days). In some other instances, organoid cells can form structures exhibiting a glomerular cluster-like feature. Such structural features can be exhibited by organoids at or after approximately 15 to 18 days of culture (e.g., approximately 15, 16, 17, or 18 days). Additionally, organoid cells can form structures exhibiting a tubular feature. Such structural features can be exhibited by organoids at or after approximately 12 to approximately 18 days of culture (e.g., approximately 12, 13, 14, 15, 16, 17, or 18 days).

[0083] Organoids prepared from enriched heterogeneous renal cell populations that express any of the markers disclosed herein or exhibit any of the structural features disclosed herein can be identified as having therapeutic potential. For example, if expression of any one or more (e.g., any one, any two, any three, any four, any five, any six, any seven, or all eight) of Hoxa11, CK8, Meis2, Thsd7a, Ptpro, CD13, LTL, and / or Cdc6 is detected, organoids prepared from enriched heterogeneous renal cell populations can be identified as having therapeutic potential. In some instances, organoids prepared from enriched heterogeneous renal cell populations can be identified as having therapeutic potential if (i) Hoxa11 and Meis2, or (ii) Hoxa11, Meis2, and CK8, or (iii) Thsd7a and Ptpro, or (iv) CD13 and LTL, or (v) Cdc6, or (vi) Hoxa11, Meis2, Thsd7a, and Ptpro, or (vii) Cdc6, Hoxa11, Meis2, and Thsd7A, or (viii) Hoxa11 and Thsd7A, or (ix) Cdc6, Hoxa11, and Thsd7A, or (x) Ptpro, or (xi) Meis2, or (xii) Thsd7A are detected. The expression of biomarkers can be detected as described herein.

[0084] Furthermore, organoids prepared from enriched heterogeneous renal cell populations can be included in pharmaceutical compositions and can be administered to patients in methods of treating kidney diseases, or can be used in methods of treating kidney diseases as described above.

[0085] Those skilled in the art will recognize, or can determine, many equivalents of the specific embodiments described herein using only conventional experiments. These equivalents are intended to be covered by the appended claims.

[0086] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference, and have the same effect as each individual publication, patent or patent application expressly and individually indicates that it is incorporated herein by reference in its entirety.

[0087] Example

[0088] Example 1 - SRC population cell expression of biomarkers related to kidney development

[0089] introduce:The general hypothesis regarding the mechanism of action of human SRC (exemplary enriched heterogeneous renal cell population) in repairing and restoring renal function in patients with nephropathy is that SRC can recapitulate aspects of fetal kidney development. In the developing kidney, signals from UB trigger postrenal mesenchymal aggregation to form renal progenitor cell caps (Plisov SY, et al., Development. 2001; 128:1045-57). CM undergoes mesenchymal-epithelial transition to form anterior tubular aggregates and renal capsules, which subsequently develop into sac-like bodies, S-like bodies, and epithelialized filtration apparatus (McMahon A., Curr Top Dev Biol. 2016;117:31-64; Thiagarajan RD, et al., PLoS One. 2011; 6:e17286, Plisov SY, et al., Development. 2001; 128:1045-57). Single-cell transcriptome analysis was performed to determine whether SRCs expressed UB, CM, and podocyte markers.

[0090] Methods - Preparation of human SRC. SRCs were isolated from human kidney biopsies obtained from the National Center for Disease Control and Prevention (NCDPC) (Philadelphia, PA; protocol number: RRON2 01 001A) or from kidney biopsies obtained from patients with type 2 diabetes (T2D) and chronic kidney disease (CKD) in a phase 2 clinical trial (NCT02836574). EasyCore was used to isolate the SRCs. TMCore renal biopsies (0.02 g) were collected using a biopsy system (Boston Scientific, Marlborough, MA), washed twice in Hanks balanced salt solution, minced, and digested for 30 min in a dispersant enzyme (Stem Cell Technologies, Canada) containing 300 U type IV collagenase (Worthington Biochemical Corporation, Lakewood, NJ) and 5 mM CaCl2 (Sigma-Aldrich, St. Louis, MO). The source biopsy cells were cultured in 35 mm tissue culture dishes (Corning / Costar, Corning, NY), and viability was determined by counting live cells at passage time. The growth rate was calculated as ln(fold increase) / time. Population doubling time was calculated as ln(2) / growth rate. Two mL suspensions of 6–75 million cells were loaded onto a density gradient (OptiPrep). TM (STEMCELL TECHNOLOGIES, Cambridge, MA), in serum-free keratinocyte culture medium without supplementation; Invitrogen, Waltham, MA), centrifuged at 800 g (without brake) for 20 minutes at room temperature. The bottom band was harvested by pipette, washed twice in sterile phosphate-buffered saline (PBS, Sigma-Aldrich), and resuspended in Dulbecco's PBS (Sigma-Aldrich).

[0091] Methods - UB, CM and podocyte markers.From publicly available kidney atlases, a preliminary set of biomarkers for UB, CM, and podocytes was generated. These atlases included the Human Fetal Kidney Atlas (Semraulab (leidenuniv.nl); last accessed 8 / 2 / 2023), The Embryonic Development, Regenerative Medicine and Stem Cell Database (LifeMap Discovery (lifemapsc.com); last accessed 8 / 2 / 2023), and The Human BioMolecular Atlas Program (HuBMAP Consortium – The HuBMAP Human BioMolecular Atlas Program; last accessed 8 / 2 / 23). Then, using fetal kidney datasets from the Kidney Cell Atlas (kidneycellatlas.org; last accessed 8 / 2 / 23), localized expression of specific biomarkers was validated, generating a refined list of anchored or classic biomarkers.

[0092] Methods - Single-cell transcriptomics study. Single-cell RNA-seq was performed by the Advanced Analytical Core Laboratory at the University of North Carolina at Chapel Hill (UNC, Chapel Hill, NC). Aliquots of SRC were prepared using acridine orange and propidium iodide (approximately 1 x 10⁻⁶ in Dulbecco's PBS). 6Cells (n=3 donors) were stained, and viability, concentration, and singleness were assessed using a LUNA-FX7 dual-fluorescence cell counter (Logos Biosystems, South Korea). Cells were then processed according to the manufacturer's instructions using a 10x Genomics Chromium Controller (Pleasanton, CA) and its Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3.1 Dual Index Kit (PN-1000268). Approximately 5,000 cells per sample were loaded onto a Chromium chip G, with a target recovery rate of 3,000 cells per sample for library preparation. Single cells, reverse transcription reagents, and gel beads coated with barcode oligos were encapsulated together in oil droplets to generate emulsion gel beads (GEMs). Reverse transcription was performed using a C1000 thermal cycler (Bio-Rad, Hercules, CA) to generate complementary DNA (cDNA) libraries tagged with cell barcodes and unique molecular indexes (UMIs). GEM was then disrupted and purified using Dynabeads MyOne SILANE beads (Invitrogen, Waltham, MA), followed by 12 cDNA amplification cycles. The amplified cDNA library was purified using SPRIselect magnetic beads (Beckman Coulter, Brea, CA) and quantified using an Agilent Bioanalyzer high-sensitivity DNA microarray (Agilent Technologies, Santa Clara, CA). Fragmentation, end repair, A-tailing, and bifacial size selection using SPRIselect beads were then performed. Illumina-compatible adaptors were ligated to the size-selected cDNA fragments. The adaptor-ligated cDNA was then purified using SPRIselect beads. Unique and identifiable indexes were added during the 12 amplification cycles. The completed sequencing library was then purified using SPRIselect beads, visualized using an Agilent Bioanalyzer high-sensitivity DNA microarray, and merged in equimolar ratios. The merged libraries were sequenced on a NextSeq 2000 instrument (Illumina, San Diego, CA) at the UNC High Throughput Sequencing Facility.Library denatured and diluted according to the standard Illumina protocol, incorporating 1% PhiX sequencing controls (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 passed through a quality filter. Demultiplexing and preliminary analysis were performed using Cell Ranger 7.1.0 with default settings. Analysis was performed using the Loupe browser from 10X Genomics (Tables 1 and 2) or Seurat (Seurat, v5.01; Tables 4 and 5).

[0093] result - Rigorous localization was performed on various publicly available fetal kidney atlases to identify typical markers, and even anchoring markers, for UB, CM, and podocytes. Preliminary analysis of marker expression was performed on SRCs prepared from kidneys obtained from NDRI, which were relatively specific for these UB, CM, and podocyte precursor populations. Genes ret, wnt11, fgf8, fgf10, grem1, etv5, cited1, eya1, hox11, ncam1, six2, osr1, and gdnf were highly specific for UB and CM and expressed by these SRCs (Table 1). Genes thsd7a, ptpro, mpp5, tppp3, lmx1b, nphs1, and nphs2 were specific for podocytes and expressed by these SRCs (Table 2).

[0094] Table 1: CM+UB markers expressed by SRC population cells

[0095]

[0096] Table 2: Podocyte markers expressed by SRC population cells

[0097]

[0098] Using these biomarkers, approximately 21.17–38.52% of the SRC population cells were found to express UB- / CM-specific genes, of which approximately 54.92–60.96% of the SRC population cells expressed podocyte biomarkers. (Table 3)

[0099] Table 3: SRC populations expressing CM+UB-specific or podocyte-specific markers

[0100]

[0101] Further SRC UB, CM, and podocyte expression analyses were performed to confirm this preliminary study. Additional analyses were conducted using SRCs prepared from renal biopsies of patients with type 2 diabetes and chronic kidney disease enrolled in a phase 2 clinical trial (NCT02836574) to investigate SRCs as potential cell therapy agents. Table 4 provides scRNA-seq expression data for the UB+CM biomarkers of these SRCs. Table 5 provides scRNA-seq expression data for the podocyte biomarkers of these SRCs.

[0102] Table 4: Percentage of SRC cells expressing CM+UB markers

[0103]

[0104] Table 5: Percentage of SRC population expressing podocyte markers

[0105]

[0106] SRC expression of these markers is consistent with the expression of markers expressed by cells involved in fetal kidney development (UB, CM, and podocytes). This gene expression data provides mechanistic insights into the repair and recovery activity of SRCs in rodent models and the stabilization of renal filtration in subjects with kidney disease in clinical trials (Stavas J, et al., KI Reports 2022; 7:1619-1629; Stavas J, et al., Blood Purif. 2023; 52:114-121).

[0107] Example 2 - Organoid expression from SRC population cells and protein expression in developing kidney cells Consistent protein biomarkers were used to identify glomerular epithelial and tubular characteristics.

[0108] introduce: To further understand the mechanism of action of SRC, SRC-derived organoids were formed in a three-dimensional (3D) culture system. The proteins expressed by the organoids and the structures formed were studied to confirm the SRC gene expression data of Example 1 and to determine the ability of SRC to form nephron-like structures in a 3D culture environment.

[0109] Methods - Organoid formation. Human SRCs were prepared as described in Example 1. The SRCs were prepared by incubating kidney cell growth medium (1x10⁻⁶ cells / mL). 6 Organoids were formed by culturing them in a rotating flask bioreactor (Corning, 3152) at 37°C / 5% CO2 for up to 18 days using a multi-magnetic stirrer (Genie; 80 rpm) in a cell / mL container.

[0110] Methods - Organoid characterization, size.Organoid diameters were measured using the EVOS M5000 imaging system (Thermo Fisher, AMF5000).

[0111] Methods - Organoid characterization and biomarkers. The fixation (4% paraformaldehyde) and blocking were followed by 3 specific markers for characterization (Table 6). Organoid protein expression was determined by staining with g / mL immunofluorescent antibody and matched isotype controls overnight at 4°C. Using a similar method, at room temperature, 1 g / mL immunofluorescent antibody was used to determine organoid protein expression. Add matched secondary antibody at g / mL and Hoechst 33342 nuclear dye for 30 minutes. Perform image analysis using Celleste analysis software (Thermo Fisher).

[0112] Table 6: Organoid characterization antibodies

[0113]

[0114] Methods - Organoid characterization and structure. Organoids cultured up to 17 days were sectioned and stained with periodic acid Schiff (PAS).

[0115] Results - In 3D culture, organoids derived from human SRCs exhibited exponential growth over an 18-day culture period. Figure 1 .

[0116] Characterization of human SRC-derived organoids by immunofluorescence antibody staining confirmed that the markers they expressed (Hoxa11 (Fig. 2B), cytokeratin 8 (Fig. 2C), and Meis2 (Fig. 2D)) were consistent with those expressed by ureteral bud + cap mesenchymal cells. The expression of markers expressed by glomerular epithelial cells (Thds7a (Fig. 3A) and Ptpro (Fig. 3B)), markers expressed by proximal tubular cells (LTL (Fig. 4A) and CD13 (Fig. 4A and 4B)), and the cell division cycle protein Cdc6 (Fig. 5B) were also detected.

[0117] Furthermore, the generation of sections from SRC-derived organoids revealed the presence of morphological features consistent with glomerular and tubular structures. Slides of SRC-derived organoids cultured for 8 days exhibited structures with a glomerular-like appearance, including PEC-like margins (Fig. 6A, arrow). In SRC-derived organoids cultured for 17 days, the organoids exhibited nephron-like features, including glomerular clusters (Fig. 6B, arrow) and tubular structures (…). Figures 7A-7B ).

[0118] Overall, the expression of characteristic markers of ureteral buds and cap mesenchyme, glomerular epithelium, and proximal tubular cells in 3D-cultured human SRC-derived organoids, as well as the acquisition of glomerular epithelial and tubular structural features, demonstrates potential repair capabilities, which underlie the therapeutic effects of SRC in CKD animal models and eGFR stabilization in patients with type 2 diabetes and CKD in clinical trials.

[0119] Example 3 - Repair effect mediated by osteopontin in selected renal cells

[0120] introduce: Osteopontin (OPN) delivers kidney repair effects. To further investigate the mechanism by which osteopontin (SRC) exhibits repair and recovery effects in a CKD model, this study aimed to determine whether OPN participates in or mediates SRC repair activity.

[0121] method: Scratch assays were performed using human SRC (National Disease Research Interchange) and renal cortical biopsy monolayers (control) in the absence / presence of OPN neutralizing antibodies. Repair activity was assessed by imaging the number of cells migrating to bridge the wound (blinded). Expression of cav1, mmp9, icam1, and cxcr4 was evaluated using qPCR. Genes were seeded into HumanBase for visualization of transcriptome function.

[0122] result: Compared with controls, SRCs showed increased migration to the wound (p<0.05), an effect reduced by the OPN antibody (p<0.05). The expression of migration markers cav1, mmp9, icam1, and CXcr4 was upregulated in SRCs (p<0.05). Both findings are consistent with those reported in WO 2011 / 143499, which is incorporated herein by reference for all purposes. Further studies using gene ontology confirmed that the OPN / SSP1-cav1-mmp9-icam1-CXcr4 axis regulates migration, adhesion, and wound healing in the kidney. See also Figure 8 .

[0123] These data suggest that SRC repair effects, such as the maintenance of renal microstructure and renal filtration function, including creatinine clearance, electrolyte balance, fluid homeostasis, urine concentration, and cystatin C metabolism, can be partially mediated by the OPN-related cascade.

[0124] Example 4 - SRC shared podocyte-parietal epithelial cell transcriptome

[0125] introduce:Effects associated with SRC administration include improved glomerular barrier function, renal filtration, and protection of renal microstructure and glomerular integrity. As a mechanism for these beneficial effects, it was investigated whether SRC shares the podocyte-parietal epithelial cell (PEC) transcriptome associated with renal repair and recovery.

[0126] method: The intersection of the podocyte-PEC gene and the directionally aligned SRC gene was input into miRNet and Cytoscape for visualization of the shared transcriptome and identification of its functional properties. As described in Example 1, human SRC (kidney from the National Center for Disease Control and Prevention) was submitted to scRNA-seq to map gene expression.

[0127] result: OSR1, 62, LHX1, FGF8, LFNG, NPHS1, NPHS2, VEGFA, CD44, and Hes1 are expressed by both podocyte-PEC and SRC cells and are involved in kidney developmental functions, including the maintenance of undifferentiated nephron precursors, the formation of renal tubules and nephrons, the maturation of the glomerular barrier, and the formation of the glomerular capillary bed. See Table 7 and... Figure 9 VEGFA appears as a pivot gene in this network. Figure 10 ).

[0128] Table 7: Podocyte-PEC gene expressed by SRC population cells

[0129]

[0130] Specifically, Hes1, a gene associated with nephron development, was found to be expressed in approximately 79% to 89% of the SRC. See Table 8 and... Figure 11 .

[0131] Table 8: SRC's hes1 expression

[0132]

[0133] The podocyte-PEC transcriptome, anchored by VEGF-A expressed by SRCs, can reenact events related to kidney development and mediate the improved glomerular integrity, renal microstructure, and renal filtration observed in CKD models.

[0134] While this disclosure has been specifically shown and described with reference to particular embodiments, some of which are preferred embodiments, those skilled in the art will understand that various changes in form and detail may be made without departing from the spirit and scope of this disclosure. Therefore, the exact summary and disclosure presented herein are not intended to be limiting.

Claims

1. A method for identifying enriched heterogeneous renal cell populations as having therapeutic potential, the method comprising: The enriched heterogeneous renal cell population was determined to express at least one of the following: (i) ureteral bud and / or cap mesenchymal (UB / CM) markers; and / or (ii) podocyte markers. The at least one UB / CM marker comprises one or more of the following: Wingless-associated integration site family member 11 (WNT11), Gremlin (GREM) 1, E26 transformation-specific (ETS) variant transcription 5 (ETV5), homeobox (HOX) 11, and neural cell adhesion molecule (NCAM) 1. The at least one podocyte marker comprises one or more of the following: thrombocytopenic leukocyte-1 domain-containing protein (THSD) 7A, protein tyrosine phosphatase receptor type O (PTPRO), matrix metalloproteinase (MPP) 5, tubulin polymerization-promoting protein (TPPP) 3, and lim homeobox transcription factor 1β (LMX1B); and If the enriched heterogeneous renal cell population is determined to express at least one of the following: (i) UB / CM markers and / or (ii) podocyte markers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

2. The method of claim 1, wherein the assay step comprises determining whether the cells of the enriched heterogeneous renal cell population express at least one of the (i) UB / CM markers, and If it is determined that the cells of the enriched heterogeneous renal cell population express at least one of the (i) UB / CM markers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

3. The method of claim 2, wherein the determination step comprises determining the percentage of cells expressing at least one (i) UB / CM marker in the enriched heterogeneous renal cell population.

4. The method of claim 2, wherein the at least one (i) UB / CM marker comprises WNT11.

5. The method of claim 3, wherein the at least one (i) UB / CM marker comprises WNT11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

6. The method of claim 2, wherein the at least one (i) UB / CM marker comprises GREM1.

7. The method of claim 3, wherein the at least one (i) UB / CM marker comprises GREM1, and If it is determined that more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

8. The method of claim 3, wherein the at least one (i) UB / CM marker comprises GREM1, and If it is determined that more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

9. The method of claim 2, wherein the at least one (i) UB / CM marker comprises ETV5.

10. The method of claim 3, wherein the at least one (i) UB / CM marker comprises ETV5, and If it is determined that more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

11. The method of claim 3, wherein the at least one (i) UB / CM marker comprises ETV5, and If it is determined that more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

12. The method of claim 2, wherein the at least one (i) UB / CM marker comprises HOX11.

13. The method of claim 3, wherein the at least one (i) UB / CM marker comprises HOX11, and If it is determined that more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

14. The method of claim 3, wherein the at least one (i) UB / CM marker comprises HOX11, and If it is determined that more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

15. The method of claim 2, wherein the at least one (i) UB / CM marker comprises NCAM1.

16. The method of claim 3, wherein the at least one (i) UB / CM marker comprises NCAM1, and If it is determined that more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

17. The method of claim 3, wherein the at least one (i) UB / CM marker comprises NCAM1, and If it is determined that more than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

18. The method of claim 2, wherein the at least one (i) UB / CM marker comprises: (a) WNT11 and GREM1; (b) WNT11 and ETV5; (c)WNT11 and HOX11; (d)WNT11 and NCAM1; (e)GREM1 and ETV5; (f)GREM1 and HOX11; (g)GREM1 and NCAM1; (h)ETV5 and HOX11; (i) ETV5 and NCAM1; or (j)HOX11 and NCAM1.

19. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: (a) WNT11 and GREM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) WNT11 and ETV5, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) WNT11 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d)WNT11 and NCAM1; and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)GREM1 and ETV5, and If it is determined that more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f)GREM1 and HOX11, and If it is determined that more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)GREM1 and NCAM1, and If it is determined that more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h)ETV5 and HOX11, and If it is determined that more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i) ETV5 and NCAM1, and If it is determined that greater than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and greater than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j) HOX11 and NCAM1, and If it is determined that more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

20. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: (a) WNT11 and GREM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) WNT11 and ETV5, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) WNT11 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d)WNT11 and NCAM1; and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, and more than 1% and at most 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)GREM1 and ETV5, and If it is determined that more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f)GREM1 and HOX11, and If it is determined that more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)GREM1 and NCAM1, and If it is determined that more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h)ETV5 and HOX11, and If it is determined that more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i) ETV5 and NCAM1, and If it is determined that greater than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and greater than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j) HOX11 and NCAM1, and If it is determined that more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and more than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

21. The method of claim 2, wherein the at least one (i) UB / CM marker comprises: (a) WNT11, GREM1 and ETV5; (b) WNT11, GREM1 and HOX11; (c)WNT11, GREM1 and NCAM1; (d)WNT11, ETV5 and HOX11; (e)WNT11, ETV5 and NCAM1; (f) WNT11, HOX11 and NCAM1; (g)GREM1, ETV5 and HOX11; (h)GREM1, ETV5 and NCAM1; (i)GREM1, HOX11, and NCAM1; or (j)ETV, HOX11 and NCAM1.

22. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: (a) WNT11, GREM1 and ETV5, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) WNT11, GREM1 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) WNT11, GREM1, and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d) WNT11, ETV5 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)WNT11, ETV5 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f) WNT11, HOX11 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)GREM1, ETV5 and HOX11, and If it is determined that more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h)GREM1, ETV5, and NCAM1, and If it is determined that more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i)GREM1, HOX11 and NCAM1, and If it is determined that greater than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, greater than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and greater than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j)ETV5, HOX11 and NCAM1, and If it is determined that more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

23. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: (a) WNT11, GREM1 and ETV5, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) WNT11, GREM1 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) WNT11, GREM1, and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, and more than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d) WNT11, ETV5 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)WNT11, ETV5 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and more than 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f) WNT11, HOX11 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and more than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)GREM1, ETV5 and HOX11, and If it is determined that more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, more than 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and more than 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h)GREM1, ETV5, and NCAM1, and If it is determined that more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i)GREM1, HOX11 and NCAM1, and If it is determined that greater than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, greater than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and greater than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j)ETV5, HOX11 and NCAM1, and If it is determined that more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and more than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

24. The method of claim 2, wherein the at least one (i) UB / CM marker comprises: (a) WNT11, GREM1, ETV5 and HOX11; (b) WNT11, GREM1, ETV5 and NCAM1; (c)WNT11, ETV5, HOX11 and NCAM1; (d) WNT11, GREM1, HOX11, and NCAM1; or (e)GREM1, ETV5, HOX11 and NCAM1.

25. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: (a) WNT11, GREM1, ETV5 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) WNT11, GREM1, ETV5 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) WNT11, ETV5, HOX11 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, more than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and more than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d) WNT11, GREM1, HOX11, and NCAM1, and If it is determined that greater than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, greater than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, greater than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and greater than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (e)GREM1, ETV5, HOX11, and NCAM1, and If it is determined that greater than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, greater than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, greater than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and greater than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

26. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: (a) WNT11, GREM1, ETV5 and HOX11, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and more than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) WNT11, GREM1, ETV5 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, more than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, and more than 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) WNT11, ETV5, HOX11 and NCAM1, and If it is determined that more than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, more than 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, more than 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and more than 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d) WNT11, GREM1, HOX11, and NCAM1, and If it is determined that greater than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, greater than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, greater than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and greater than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (e)GREM1, ETV5, HOX11, and NCAM1, and If it is determined that more than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, more than 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, more than 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and more than 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

27. The method of claim 2, wherein the at least one (i) UB / CM marker comprises: WNT11, GREM1, ETV5, HOX11 and NCAM1.

28. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: WNT11, GREM1, ETV5, HOX11, and NCAM1, and If it is determined that greater than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, greater than 0% and at most about 5% of the enriched heterogeneous renal cell population expresses GREM1, greater than about 2% and at most about 25% of the enriched heterogeneous renal cell population expresses ETV5, greater than about 2% and at most about 20% of the enriched heterogeneous renal cell population expresses HOX11, and greater than about 0.25% and at most about 2.5% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

29. The method of claim 3, wherein the at least one (i) UB / CM marker comprises: WNT11, GREM1, ETV5, HOX11, and NCAM1, and If it is determined that greater than 0% and at most about 1% of the enriched heterogeneous renal cell population expresses WNT11, greater than 5% and at most about 35% of the enriched heterogeneous renal cell population expresses GREM1, greater than about 50% and at most about 70% of the enriched heterogeneous renal cell population expresses ETV5, greater than about 15% and at most about 30% of the enriched heterogeneous renal cell population expresses HOX11, and greater than about 1% and at most about 7% of the enriched heterogeneous renal cell population expresses NCAM1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

30. The method of claim 1, wherein the assay step comprises determining whether cells in the enriched heterogeneous renal cell population express at least one of the (ii) podocyte markers, and If it is determined that the cells of the enriched heterogeneous renal cell population express at least one of the (ii) podocyte markers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

31. The method of any one of claims 2-30, wherein the determining step further comprises determining whether the cells of the enriched heterogeneous renal cell population express at least one of the (ii) podocyte markers, and If it is further determined that the cells of the enriched heterogeneous renal cell population express at least one of the (ii) podocyte markers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

32. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises THSD7A.

33. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises PTPRO.

34. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises MPP5.

35. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises TPPP3.

36. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises LMX1B.

37. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A and PTPRO; (b) THSD7A and MPP5; (c)THSD7A and TPPP3; (d)THSD7A and LMX1B; (e)PTPRO and MPP5; (f) PTPRO and TPPP3; (g)PTPRO and LMX1B; (h)MPP5 and TPPP3; (i) MPP5 and LMX1B; or (j)TPPP3 and LMX1B.

38. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A, PTPRO, and MPP5; (b) THSD7A, PTPRO, and TPPP3; (c)THSD7A, PTPRO, and LMX1B; (d)THSD7A, MPP5 and TPPP3; (e)THSD7A, MPP5 and LMX1B; (f)THSD7A, TPPP3 and LMX1B; (g)PTPRO, MPP5 and TPPP3; (h)PTPRO, MPP5 and LMX1B; (i) PTPRO, TPPP3, and LMX1B; or (j) MPP5, TPPP3 and LMX1B.

39. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A, PTPRO, MPP5 and TPPP3; (b) THSD7A, PTPRO, MPP5, and LMX1B; (c)THSD7A, MPP5, TPPP3 and LMX1B; (d) THSD7A, PTPRO, TPPP3, and LMX1B; or (e)GREM1, MPP5, TPPP3 and LMX1B.

40. The method of claim 30 or 31, wherein the at least one (ii) podocyte marker comprises THSD7A, PTPRO, MPP5, TPPP3 and LMX1B.

41. The method of claim 30 or 31, wherein the determination step comprises determining the percentage of cells expressing at least one of the (ii) podocyte markers in the enriched heterogeneous renal cell population.

42. The method of claim 41, wherein the at least one (ii) podocyte marker comprises THSD7A, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

43. The method of claim 41, wherein the at least one (ii) podocyte marker comprises THSD7A, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

44. The method of claim 41, wherein the at least one (ii) podocyte marker comprises PTPRO, and If it is determined that more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

45. The method of claim 41, wherein the at least one (ii) podocyte marker comprises PTPRO, and If it is determined that more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

46. ​​The method of claim 41, wherein the at least one (ii) podocyte marker comprises MPP5, and If it is determined that more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

47. The method of claim 41, wherein the at least one (ii) podocyte marker comprises MPP5, and If it is determined that more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

48. The method of claim 41, wherein the at least one (ii) podocyte marker comprises TPPP3, and If it is determined that more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

49. The method of claim 41, wherein the at least one (ii) podocyte marker comprises LMX1B, and If it is determined that more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population express LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

50. The method of claim 41, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A and PTPRO, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, and more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) THSD7A and MPP5, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, and more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) THSD7A and TPPP3, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d)THSD7A and LMX1B, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)PTPRO and MPP5, and If it is determined that more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f) PTPRO and TPPP3, and If it is determined that more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)PTPRO and LMX1B, and If it is determined that more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h) MPP5 and TPPP3, and If it is determined that more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i) MPP5 and LMX1B, and If it is determined that greater than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j)TPPP3 and LMX1B, and If it is determined that more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

51. The method of claim 41, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A and PTPRO, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, and more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) THSD7A and MPP5, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, and more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) THSD7A and TPPP3, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d)THSD7A and LMX1B, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)PTPRO and MPP5, and If it is determined that more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f) PTPRO and TPPP3, and If it is determined that more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)PTPRO and LMX1B, and If it is determined that more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h) MPP5 and TPPP3, and If it is determined that more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i) MPP5 and LMX1B, and If it is determined that greater than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j)TPPP3 and LMX1B, and If it is determined that more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

52. The method of claim 41, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A, PTPRO, and MPP5, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) THSD7A, PTPRO, and TPPP3, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) THSD7A, PTPRO, and LMX1B, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d)THSD7A, MPP5, and TPPP3; and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)THSD7A, MPP5, and LMX1B, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f) THSD7A, TPPP3, and LMX1B, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)PTPRO, MPP5 and TPPP3, and If it is determined that more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h)PTPRO, MPP5, and LMX1B, and If it is determined that more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i) PTPRO, TPPP3 and LMX1B, and If it is determined that greater than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j) MPP5, TPPP3 and LMX1B, and If it is determined that more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

53. The method of claim 41, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A, PTPRO, and MPP5, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) THSD7A, PTPRO, and TPPP3, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) THSD7A, PTPRO, and LMX1B, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d)THSD7A, MPP5, and TPPP3; and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (e)THSD7A, MPP5, and LMX1B, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (f) THSD7A, TPPP3, and LMX1B, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (g)PTPRO, MPP5 and TPPP3, and If it is determined that more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (h)PTPRO, MPP5, and LMX1B, and If it is determined that more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (i) PTPRO, TPPP3 and LMX1B, and If it is determined that greater than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (j) MPP5, TPPP3 and LMX1B, and If it is determined that more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

54. The method of claim 41, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A, PTPRO, MPP5, and TPPP3, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) THSD7A, PTPRO, MPP5, and LMX1B, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) THSD7A, MPP5, TPPP3 and LMX1B, and If it is determined that more than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than about 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d) THSD7A, PTPRO, TPPP3, and LMX1B, and If it is determined that greater than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, greater than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (e)PTPRO, MPP5, TPPP3 and LMX1B, and If it is determined that greater than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, greater than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

55. The method of claim 41, wherein the at least one (ii) podocyte marker comprises: (a) THSD7A, PTPRO, MPP5, and TPPP3, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (b) THSD7A, PTPRO, MPP5, and LMX1B, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (c) THSD7A, MPP5, TPPP3 and LMX1B, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. (d) THSD7A, PTPRO, TPPP3, and LMX1B, and If it is determined that greater than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, greater than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential; or (e)PTPRO, MPP5, TPPP3 and LMX1B, and If it is determined that more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

56. The method of claim 41, wherein the at least one (ii) podocyte marker comprises THSD7A, PTPRO, MPP5, TPPP3, and LMX1B, and If it is determined that greater than about 1.5% and at most about 6% of the enriched heterogeneous renal cell population expresses THSD7A, greater than about 0.1% and at most about 3% of the enriched heterogeneous renal cell population expresses PTPRO, greater than about 40% and at most about 70% of the enriched heterogeneous renal cell population expresses MPP5, greater than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and greater than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

57. The method of claim 41, wherein the at least one (ii) podocyte marker comprises THSD7A, PTPRO, MPP5, TPPP3, and LMX1B, and If it is determined that more than about 10% and at most about 30% of the enriched heterogeneous renal cell population expresses THSD7A, more than about 2.5% and at most about 10% of the enriched heterogeneous renal cell population expresses PTPRO, more than about 80% and at most about 95% of the enriched heterogeneous renal cell population expresses MPP5, more than about 0.1% and at most about 0.6% of the enriched heterogeneous renal cell population expresses TPPP3, and more than 0% and at most about 0.4% of the enriched heterogeneous renal cell population expresses LMX1B, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

58. The method of any one of the preceding claims, further comprising detecting at least one additional marker, The at least one additional marker is an additional UB / CM marker. The aforementioned additional biomarker comprises one or more of the following: transfection rearrangement (RET), fibroblast growth factor (FGF) 8, FGF 10, cbp / P300 interaction transactivator (CITED) 1, eye loss (EYA) 1, sine oculis homeobox homologue 2 (SIX2), odd-skipped associated protein 1 (OSR1), and / or glial cell-derived neurotrophic factor (GDNF). If the enriched heterogeneous renal cell population is further determined to express at least one of the additional markers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

59. The method of any one of the preceding claims, wherein the assay step further comprises determining whether the cells of the enriched heterogeneous renal cell population express at least one podocyte-PEC marker, and If it is further determined that the enriched heterogeneous renal cell population expresses at least one of the podocyte-PEC markers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential. The at least one podocyte-PEC marker mentioned above includes hes1.

60. The method of claim 59, wherein determining whether cells in the enriched heterogeneous renal cell population express the at least one podocyte-PEC marker comprises determining the percentage of cells in the enriched heterogeneous renal cell population expressing the at least one podocyte-PEC marker.

61. The method of claim 60, wherein if it is determined that more than about 70% of the enriched heterogeneous renal cell population expresses hes1, the enriched heterogeneous renal cell population is identified as having therapeutic potential.

62. The method of any one of the preceding claims, further comprising: The study determined whether the enriched heterogeneous renal cell population expressed transforming growth factor β2 (TGFβ2). If it is determined that the enriched heterogeneous renal cell population expresses TGFβ2, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

63. The method of claim 62, wherein the determination comprises determining the percentage of cells expressing TGFβ2 in the enriched heterogeneous renal cell population, and If it is determined that more than about 50% of the cells in the enriched heterogeneous renal cell population express TGFβ2, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

64. The method of claim 62, wherein the determination includes determining whether the cells of the enriched heterogeneous renal cell population secrete TGFβ2.

65. The method of claim 64, wherein determining whether the cells of the enriched heterogeneous renal cell population secrete TGFβ2 further comprises determining the amount of TGFβ2 secreted by said cells. If it is determined that each 1,000,000 enriched heterogeneous renal cell population secretes at least about 1.0 ng TGFβ2, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

66. A method for identifying enriched heterogeneous populations of renal cells as having therapeutic potential, said method comprising: The percentage of kidney cells expressing ureteral bud (UB), cap mesenchymal (CM), and podocyte characteristic genes in the population was determined; and The population is identified as having therapeutic potential if the following conditions are met: (i) Approximately 20% to 40% of the renal cells in the population express UB and CM cell characteristic genes; and (ii) Approximately 50% to 65% of the kidney cells in the population express podocyte-specific genes; The population was prepared from non-fetal kidney tissue.

67. The method of claim 66, wherein the UB and CM cell characteristic genes comprise: Wingless-associated integration site family member 11 (WNT11), Gremlin (GREM) 1, E26 transformation-specific (ETS) variant transcription 5 (ETV5), homeobox (HOX) 11, neural cell adhesion molecule (NCAM) 1, transfection-induced rearrangement (RET), fibroblast growth factor 8 (FGF8), FGF10, cbp / P300 interaction transactivator (CITED) 1, eye absence (EYA) 1, sine oculis homeobox homologue 2 (SIX2), odd-skipped associated 1 (OSR1), and glial cell-derived neurotrophic factor (GDNF).

68. The method of claim 66, wherein the podocyte characteristic gene comprises matrix metalloproteinase (MPP) 5.

69. The method of claim 67, wherein the podocyte characteristic gene comprises matrix metalloproteinase (MPP) 5.

70. The method of claim 68 or 69, wherein the podocyte characteristic gene further comprises platelet-reactive protein type 1 domain protein (THSD) 7A, protein tyrosine phosphatase receptor type O (PTPRO), tubulin polymerization promoting protein (TPPP) 3, lim homeobox transcription factor 1β (LMX1B), and nephrotic protein (NPHS1).

71. The method of any one of claims 66, 67 or 70, wherein the UB and CM cell characteristic genes consist of WNT11, GREM1, ETV5, HOX11, NCAM1, RET, FGF8, FGF10, CITED1, EYA1, SIX2, OSR1 and GDNF.

72. The method of any one of claims 66, 67 or 70, wherein the podocyte characteristic gene comprises THSD7A, PTPRI, MMP5, TPPP3, LIMX1B and NPHS1.

73. An enriched heterogeneous population of kidney cells, identified according to the method of any one of the preceding claims.

74. A pharmaceutical composition comprising the enriched heterogeneous renal cell population of claim 73.

75. A method for treating kidney disease in patients with this need, said method comprising: Administer a therapeutically effective amount of the pharmaceutical composition of claim 74.

76. Use of the pharmaceutical composition of claim 74 in the preparation of a medicament for treating kidney disease.

77. The enriched heterogeneous renal cell population of claim 73, wherein the population is prepared by a method including a step of centrifugation using a density medium.

78. The enriched heterogeneous renal cell population of claim 77, wherein the population comprises cells with a buoyancy density greater than about 1.04 g / mL.

Images