Selected renal cell populations, features and uses thereof

Abstract

A method of identifying an enriched heterogeneous kidney cell population having therapeutic potential, an enriched heterogeneous kidney cell population having therapeutic potential and uses thereof.

Description

Background Art

[0001] Chronic kidney disease (CKD) is at or has reached pandemic proportions (Thurlow JS, et al., Am J Nephrol 52(2021):98-107; Murphy D, et al., Intern Med 65(2016):473-481; Hill NR, et al., PLoS One 11:e0158765. doi: 10.1371 / journal.pone.0158765). Incidence data from the United States to Europe show that approximately 10% of the general population has stage 1-3 CKD (ERA, 2009; USRDS, 2011; Jha et al., Lancet. 382(2013):260-72). Between 1996 and 2006, in the United States alone, CKD increased by more than 33% (U.S. Renal Data System. Costs of CKD and ESRD. Minneapolis, MN, 2007). At the same time, the management of CKD remains a challenge for nephrologists. Most standard care treatments for CKD are small molecules that target biochemical pathways in the kidney to affect single or related comorbidities. However, these treatments do not affect the glomerular and tubulointerstitial dysfunction of the underlying CKD kidney. Ultimately, in patients progressing to end-stage renal disease (ESRD), renal replacement therapy (RRT; dialysis or transplantation) is required to survive. There is still a lack of treatment modalities that directly address the biology of diseased kidney tissue. Such treatment modalities may prevent or even reverse the progression of CKD and reduce or avoid the need for RRT.

[0002] Selected renal cells (SRC), a heterogeneous population of renal cells rich in renal epithelial cells, are currently being advanced as an autologous cell-based therapy for the treatment of CKD (Stavas J, et al., Am J Nephrol 53(2022):50-58.; Stavas J, et al., KI Reports, to be published). SRC are derived from donor kidneys and are mainly a mixture of proximal tubular epithelial cells (Presnell SC, et al, Tissue Eng Part C Methods 17(2011):261-73). The results of a phase II clinical trial in a diabetic nephropathy cohort showed that SRC administration was safe and was accompanied by an improvement in renal function (Stavas J, et al., KI Reports, to be published).

[0003] There is a need in the art to identify heterogeneous populations of renal cells, such as SRC, that can improve renal function in patients with CKD and reduce or avoid the need for RRT. SUMMARY OF THE INVENTION

[0005] The present disclosure describes methods for identifying enriched heterogeneous populations of renal cells with therapeutic potential. In the method, it is determined whether the cells in the enriched heterogeneous population of renal cells express (i) NEPH1 and (ii) one or more markers, where the one or more markers can be nephrin (NPHS1), podocin (NPHS2), sine oculis homeobox homolog 2 (SIX2), odd skipped related 1 (OSR1), LIM homeobox protein 1 (LHX1), fibroblast growth factor 8 (FGF8), receptor for activated C kinase 1 (RACK1), or rearranged during transfection (RET). If the cells in the enriched heterogeneous population of renal cells express (i) NEPH1 and (ii) one or more markers, the enriched heterogeneous population of renal cells is identified as having therapeutic potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0006] Figure 1A-1B : Description of the output of co-expression interactome analysis by STRING( Figure 1A ) and Genemania( Figure 1B ) in human tissues of eight nodes expressed on an enriched heterogeneous population of renal cells (e.g., human selected renal cells (SRC)); eight nodes = Six2, Osr1, Lhx1, Ret, Fgf8, Rack1, Nphs1, Nphs2, ret, and rack1 / gnb211; co-expression of nodes ret and rack1 / gnb211 with nodes Six2, Osr1, Lhx1, Ret, Fgf8, Rack1, Nphs1, and Nphs2 was not found in human tissues in the interactome analysis.

[0007] Figure 2A-2H : Violin plots generated from the Human Fetal Kidney Atlas are provided, reporting six2( Figure 2A ), osr1( Figure 2B ), lhx1( Figure 2C ), ret( Figure 2D ), fgf8( Figure 2E ), rack1( Figure 2F ), nphs1( Figure 2G ), and nphs2( Figure 2H) expression. Renal cell types: NPCa, nephron progenitor cell subtype a; NPCb, nephron progenitor cell subtype b; NPCc, nephron progenitor cell subtype c; NPCd, nephron progenitor cell subtype d; PTA, pretubular aggregate cells, RVCSBa, renal vesicle / comma-shaped body cell subtype a; RVCSBb, renal vesicle / comma-shaped body cell subtype b; SSBpr, proximal S-shaped body cells; SSBm / d, medial / distal S-shaped body cells, SSBpod, S-shaped body cell podocyte progenitor; CnT, connecting tubule cells; DTLH, distal renal tubule / loop of Henle cells; ErPrT, early proximal renal tubule cells; Pod, podocytes; UBCD, ureteric bud / collecting duct cells, IPC, interstitial progenitor cells; ICa, interstitial cell subtype a, ICb interstitial cell subtype b; Mes, mesangial cells; End, endothelial cells; Leu, leukocytes; Prolif, proliferating cells.

[0008] Figure 3 : Provided is a human SRC interactome constructed by mapping the interactions between eight nodes co-expressed with human SRC. In addition to the eight nodes co-expressed with SRC, the SRC interactome may also include other nodes, such as Kirre-like nephrin family adhesion molecule 1 (kirrel 1, whose expression product is NEPH1).

[0009] Figure 4 : Provided is a gene ontology (GO) biological process map of 40 pathways related to human SRC generated using ShinyGO. Notably, these pathways are mainly involved in neonephrogenesis.

[0010] Figure 5A-5B : Provided is a biological process gene network analysis showing the correlation of the human SRC interactome with angiogenesis (network strength = 0.45) and Na + transport (network strength = 0.74). The biological process gene network analysis was performed using Human Base.

[0011] Figure 6 : Shows the uniform manifold approximation and projection (UMAP) of SRC population cells expressing kirrel1. Dark gray dots represent cells expressing kirrel1. Light gray dots represent cells not expressing kirrel1. DETAILED DESCRIPTION OF THE INVENTION

[0013] Described herein are methods for identifying enriched heterogeneous renal cell populations with therapeutic potential, including compositions identified as enriched heterogeneous renal cell populations with therapeutic potential, and methods and uses of enriched heterogeneous renal cell populations with therapeutic potential.

[0014] In a method for identifying an enriched heterogeneous renal cell population having therapeutic potential, the therapeutic potential of the enriched heterogeneous renal cell population can be for treating kidney disease, renal tubular transport defects, or glomerular filtration defects.

[0015] The therapeutic potential of the enriched heterogeneous renal cell population identified by this method can be for renal function restoration, renal function stabilization, renal function improvement, reduction of renal fibrosis, reduction of renal inflammation, induction of renal tubule formation in the kidney, induction of nephrogenesis in the kidney, or induction of glomerulus formation in the kidney in a patient in need of such treatment. If the enriched heterogeneous renal cell population is identified as having therapeutic potential, the enriched heterogeneous renal cell population can restore mineral balance or alleviate anemia in a patient in need of such treatment. If the enriched heterogeneous renal cell population is identified as having therapeutic potential, it can delay or prevent the need for dialysis, or it can delay or prevent the need for kidney transplantation in a patient in need of treatment for kidney disease.

[0016] In a method for identifying an enriched heterogeneous renal cell population having therapeutic potential, it can be determined whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 (also known as KIRREL1, KIRREL, IRRE-like kin (Drosophila), Kin of IRRE-like protein 1, kirre-like nephropathy protein family adhesion molecule 1, NPHS23) and (ii) one or more markers. The one or more markers can be or can include any one or more of the following: nephrin, podocin, homeobox protein sine oculis homeobox homolog 2 (SIX2), odd-skipped related 1 (OSR1), lim homeobox protein 1 (LHX1), fibroblast growth factor 8 (FGF8), receptor for activated C kinase 1 (RACK1), or rearranged during transfection (RET). The one or more markers can be or can include any one, any two, any three, any four, any five, any six, any seven, or all eight of nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1.

[0017] In a method for identifying an enriched heterogeneous renal cell population with therapeutic potential, it can be determined whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) at least two markers among nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1. If the expression of at least two markers is determined, the at least two markers can be or can include (a) SIX2 and OSR1; (b) SIX2 and LHX1; (c) SIX2 and RET; (d) SIX2 and FGF8; (e) OSR1 and LHX1; (f) OSR1 and RET; (g) OSR1 and FGF8; (h) LHX1 and RET; (i) LHX1 and FGF8; (j) RET and FGF8; (k) nephrin and SIX2; (l) nephrin and OSR1; (m) nephrin and LHX1, (n) nephrin and RET, (o) nephrin and FGF8; (p) podocin and nephrin; (q) podocin and SIX2; (r) podocin and OSR1; (s) podocin and LHX1; (t) podocin and RET; (u) podocin and FGF8; (v) RACK1 and nephrin; (w) RACK1 and podocin; (x) RACK1 and SIX2; (y) RACK1 and OSR1; (z) RACK1 and LHX1; (aa) RACK1 and RET; or (bb) RACK1 and FGF8.

[0018] In a method for identifying an enriched heterogeneous renal cell population having therapeutic potential, it can be determined whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) at least three markers among nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1. If the expression of at least three markers is determined, the at least three markers can be or can include (a) SIX2, OSR1, and LHX1; (b) SIX2, OSR1, and RET; (c) SIX2, OSR1, and FGF8; (d) SIX2, LHX1, and RET; (e) SIX2, LHX1, and FGF8; (f) SIX2, RET, and FGF8; (g) OSR1, LHX1, and RET; (h) OSR1, LHX1, and FGF8; (i) OSR1, RET, and FGF8; (j) LHX1, RET, and FGF8; (k) nephrin, SIX2, and OSR; (l) nephrin, SIX2, and LHX1; (m) nephrin, SIX2, and RET; (n) nephrin, SIX2, and FGF8; (o) nephrin, OSR, and LHX1; (p) nephrin, OSR, and RET; (q) nephrin, OSR, and FGF8; (r) nephrin, LHX1, and RET; (s) nephrin, LHX1, and FGF8; (t) nephrin, RET, and FGF8; (u) podocin, SIX2, and OSR; (v) podocin, SIX2, and LHX1; (w) podocin, SIX2, and RET; (x) podocin, SIX2, and FGF8; (y) podocin, OSR, and LHX1; (z) podocin, OSR, and RET; (aa) podocin, OSR, and FGF8; (bb) podocin, LHX1, and RET; (cc) podocin, LHX1, and FGF8; (dd) podocin, RET, and FGF8; (ee) RACK1, SIX2, and OSR; (ff) RACK1, SIX2, and LHX1; (gg) RACK1, SIX2, and RET; (hh) RACK1, SIX2, and FGF8; (ii) RACK1, OSR, and LHX1; (jj) RACK1, OSR, and RET; (kk) RACK1, OSR, and FGF8; (ll) RACK1, LHX1, and RET; (mm) RACK1, LHX1, and FGF8; (nn) RACK1, RET, and FGF8; (oo) nephrin, podocin, and RACK1; (pp) nephrin, podocin, and SIX2; (qq) nephrin, podocin, and OSR; (rr) nephrin, podocin, and RET; (ss) nephrin, podocin, and LHX1; (tt) nephrin, podocin, and FGF8; (uu) nephrin, RACK1, and SIX2; (vv) nephrin, RACK1, and OSR;(ww) Nephrin, RACK1, and RET; (xx) Nephrin, RACK1, and LHX1; (yy) Nephrin, RACK1, and FGF8; (zz) Podocin, RACK1, and SIX2; (aaa) Podocin, RACK1, and OSR; (bbb) Podocin, RACK1, and RET; (ccc) Podocin, RACK1, and LHX1; or (ddd) Podocin, RACK1, and FGF8.;

[0019] In a method for identifying an enriched heterogeneous renal cell population with therapeutic potential, it can be determined whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) at least four markers among nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1. If the expression of at least four markers is determined, the at least four markers can be or can include (a) SIX2, OSR1, LHX1, and RET; (b) SIX2, OSR1, LHX1, and FGF8; (c) SIX2, LHX1, RET, and FGF8; (d) SIX2, OSR1, RET, and FGF8; (e) OSR1, LHX1, RET, and FGF8; (f) nephrin, SIX2, OSR1, and LHX1; (g) nephrin, SIX2, OSR1, and RET; (h) nephrin, SIX2, OSR1, and FGF8; (i) nephrin, SIX2, LHX1, and RET; (j) nephrin, SIX2, LHX1, and FGF8; (k) nephrin, SIX2, RET, and FGF8; (l) nephrin, OSR1, LHX1, and FGF8; (m) nephrin, OSR1, LHX1, and RET; (n) nephrin, OSR1, RET, and FGF8; (o) nephrin, LHX1, RET, and FGF8; (p) nephrin, SIX2, OSR1, and LHX1; (q) podocin, SIX2, OSR1, and RET; (r) podocin, SIX2, OSR1, and FGF8; (s) podocin, SIX2, LHX1, and RET; (t) podocin, SIX2, LHX1, and FGF8; (u) podocin, SIX2, RET, and FGF8; (v) podocin, OSR1, LHX1, and FGF8; (w) podocin, OSR1, LHX1, and RET; (x) podocin, OSR1, RET, and FGF8; (y) podocin, LHX1, RET, and FGF8; (z) RACK1, SIX2, OSR1, and FGF8; (aa) RACK1, SIX2, LHX1, and RET; (bb) RACK1, SIX2, LHX1, and FGF8; (cc) RACK1, SIX2, RET, and FGF8; (dd) RACK1, OSR1, LHX1, and FGF8; (ee) RACK1, OSR1, LHX1, and RET; (ff) RACK1, OSR1, RET, and FGF8; (gg) RACK1, LHX1, RET, and FGF8; (hh) nephrin, podocin, SIX2, and OSR1; (ii) nephrin, podocin, SIX2, and LHX1; (jj) nephrin, podocin, SIX2, and RET; (kk) nephrin, podocin, SIX2, and FGF8; (ll) nephrin, podocin, OSR1, and LHX1;(mm) Nephrin, podocin, OSR1 and RET; (nn) Nephrin, podocin, OSR1 and FGF8; (oo) Nephrin, podocin, LHX1 and RET; (pp) Nephrin, podocin, LHX1 and FGF8; (qq) Nephrin, podocin, RET and FGF8; (rr) RACK1, podocin, SIX2 and OSR1; (ss) RACK1, podocin, SIX2 and LHX1; (tt) RACK1, podocin, SIX2 and RET; (uu) RACK1, podocin, SIX2 and FGF8; (vv) RACK1, podocin, OSR1 and LHX1; (ww) RACK1, podocin, OSR1 and RET; (xx) RACK1, podocin, OSR1 and FGF8; (yy) RACK1, podocin, LHX1 and RET; (zz) RACK1, podocin, LHX1 and FGF8; (aaa) RACK1, podocin, RET and FGF8; (bbb) RACK1, nephrin, SIX2 and OSR1; (ccc) RACK1, nephrin, SIX2 and LHX1; (ddd) RACK1, nephrin, SIX2 and RET; (eee) RACK1, nephrin, SIX2 and FGF8; (fff) RACK1, nephrin, OSR1 and LHX1; (ggg) RACK1, nephrin, OSR1 and RET; (hhh) RACK1, nephrin, OSR1 and FGF8; (iii) RACK1, nephrin, LHX1 and RET; (jjj) RACK1, nephrin, LHX1 and FGF8; (kkk) RACK1, nephrin, RET and FGF8; (lll) RACK1, nephrin, podocin and SIX2; (mmm) RACK1, nephrin, podocin and RET; (nnn) RACK1, nephrin, podocin and LHX1; (ooo) RACK, nephrin, podocin and OSR1; or (ppp) RACK, nephrin, podocin and FGF8.;

[0020] In a method for identifying an enriched heterogeneous renal cell population having therapeutic potential, it can be determined whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) at least five markers out of nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1. If the expression of at least five markers is determined, the at least five markers can be or can include (a) SIX2, OSR1, LHX1, RET, and FGF8; (b) nephrin, SIX2, OSR1, LHX1, and RET; (c) nephrin, SIX2, OSR1, LHX1, and FGF8; (d) nephrin, SIX2, LHX1, RET, and FGF8; (e) nephrin, SIX2, OSR1, RET, and FGF8; (f) nephrin, OSR1, LHX1, RET, and FGF8; (g) podocin, SIX2, OSR1, LHX1, and RET; (h) podocin, SIX2, OSR1, LHX1, and FGF8; (i) podocin, SIX2, LHX1, RET, and FGF8; (j) podocin, SIX2, OSR1, RET, and FGF8; (k) podocin, OSR1, LHX1, RET, and FGF8; (l) RACK1, SIX2, OSR1, LHX1, and RET; (m) RACK1, SIX2, OSR1, LHX1, and FGF8; (n) RACK1, SIX2, LHX1, RET, and FGF8; (o) RACK1, SIX2, OSR1, RET, and FGF8; (p) RACK1, OSR1, LHX1, RET, and FGF8; (q) nephrin, podocin, SIX2, OSR1, and LHX1; (r) nephrin, podocin, SIX2, OSR1, and RET; (s) nephrin, podocin, SIX2, OSR1, and FGF8; (t) nephrin, podocin, SIX2, LHX1, and RET; (u) nephrin, podocin, SIX2, LHX1, and FGF8; (v) nephrin, podocin, SIX2, RET, and FGF8; (w) nephrin, podocin, OSR1, LHX1, and RET; (x) nephrin, podocin, OSR1, LHX1, and FGF8; (y) nephrin, podocin, OSR1, RET, and FGF8; (z) nephrin, podocin, LHX1, RET, and FGF8; (aa) nephrin, RACK1, SIX2, OSR1, and LHX1; (bb) nephrin, RACK1, SIX2, OSR1, and RET; (cc) nephrin, RACK1, SIX2, OSR1, and FGF8; (dd) nephrin, RACK1, SIX2, LHX1, and RET; (ee) nephrin, RACK1, SIX2, LHX1, and FGF8;(ff) Nephrin, RACK1, SIX2, RET, and FGF8; (gg) Nephrin, RACK1, OSR1, LHX1, and RET; (hh) Nephrin, RACK1, OSR1, LHX1, and FGF8; (ii) Nephrin, RACK1, OSR1, RET, and FGF8; (jj) Nephrin, RACK1, LHX1, RET, and FGF8; (kk) Podocin, RACK1, SIX2, OSR1, and LHX1; (ll) Podocin, RACK1, SIX2, OSR1, and RET; (mm) Podocin, RACK1, SIX2, OSR1, and FGF8; (nn) Podocin, RACK1, SIX2, LHX1, and RET; (oo) Podocin, RACK1, SIX2, LHX1, and FGF8; (pp) Podocin, RACK1, SIX2, RET, and FGF8; (qq) Podocin, RACK1, OSR1, LHX1, and RET; (rr) Podocin, RACK1, OSR1, LHX1, and FGF8; (ss) Podocin, RACK1, OSR1, RET, and FGF8; (tt) Podocin, RACK1, LHX1, RET, and FGF8; (uu) Nephrin, Podocin, RACK1, SIX2, and OSR1; (vv) Nephrin, Podocin, RACK1, SIX2, and LHX1; (ww) Nephrin, Podocin, RACK1, SIX2, and RET; (xx) Nephrin, Podocin, RACK1, SIX2, and FGF8; (yy) Nephrin, Podocin, RACK1, OSR1, and LHX1; (zz) Nephrin, Podocin, RACK1, OSR1, and RET; (aaa) Nephrin, Podocin, RACK1, OSR1, and FGF8; (bbb) Nephrin, Podocin, RACK1, LHX1, and RET; (ccc) Nephrin, Podocin, RACK1, LHX1, and FGF8; or (ddd) Nephrin, Podocin, RACK1, RET, and FGF8.;

[0021] In a method of identifying an enriched heterogeneous renal cell population having therapeutic potential, it can be determined whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) at least six markers among nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1.If the expression of at least six markers is determined, the at least six markers may be or may include (a) nephrin, SIX2, OSR1, LHX1, RET, and FGF8; (b) podocin, SIX2, OSR1, LHX1, RET, and FGF8; (c) RACK1, SIX2, OSR1, LHX1, RET, and FGF8; (d) nephrin, podocin, SIX2, OSR1, LHX1, and RET; (e) nephrin, podocin, SIX2, OSR1, LHX1, and FGF8; (f) nephrin, podocin, SIX2, LHX1, RET, and FGF8; (g) nephrin, podocin, SIX2, OSR1, RET, and FGF8; (h) nephrin, podocin, OSR1, LHX1, RET, and FGF8; (i) nephrin, RACK1, SIX2, OSR1, LHX1, and RET; (j) nephrin, RACK1, SIX2, OSR1, LHX1, and FGF8; (k) nephrin, RACK1, SIX2, LHX1, RET, and FGF8; (l) nephrin, RACK1, SIX2, OSR1, RET, and FGF8; (m) nephrin, RACK1, OSR1, LHX1, RET, and FGF8; (n) podocin, RACK1, SIX2, OSR1, LHX1, and RET; (o) podocin, RACK1, SIX2, OSR1, LHX1, and FGF8; (p) podocin, RACK1, SIX2, LHX1, RET, and FGF8; (q) podocin, RACK1, SIX2, OSR1, RET, and FGF8; (r) podocin, RACK1, OSR1, LHX1, RET, and FGF8; (s) nephrin, podocin, RACK1, SIX2, OSR1, and LHX1; (t) nephrin, podocin, RACK1, SIX2, OSR1, and RET; (u) nephrin, podocin, RACK1, SIX2, OSR1, and FGF8; (v) nephrin, podocin, RACK1, SIX2, LHX1, and RET; (w) nephrin, podocin, RACK1, SIX2, LHX1, and FGF8; (x) nephrin, podocin, RACK1, SIX2, RET, and FGF8; (y) nephrin, podocin, RACK1, OSR1, LHX1, and RET; (z) nephrin, podocin, RACK1, OSR1, LHX1, and FGF8; (aa) nephrin, podocin, RACK1, OSR1, RET, and FGF8; or (bb) nephrin, podocin, RACK1, LHX1, RET, and FGF8.

[0022] In a method of identifying an enriched heterogeneous renal cell population having therapeutic potential, it can be determined whether cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) at least seven of the markers nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1. If the expression of at least seven markers is determined, the at least seven markers can be or can include (a) podocin, RACK1, SIX2, OSR1, LHX1, RET, and FGF8; (b) nephrin, RACK1, SIX2, OSR1, LHX1, RET, and FGF8; (c) nephrin, podocin, SIX2, OSR1, LHX1, RET, and FGF8; (d) nephrin, podocin, RACK1, OSR1, LHX1, RET, and FGF8; (e) nephrin, podocin, RACK1, SIX2, LHX1, RET, and FGF8; (f) nephrin, podocin, RACK1, SIX2, OSR1, RET, and FGF8; (g) nephrin, podocin, RACK1, SIX2, OSR1, LHX1, and FGF8; or (h) nephrin, podocin, RACK1, SIX2, OSR1, LHX1, and RET.

[0023] In a method of identifying an enriched heterogeneous renal cell population having therapeutic potential, it can be determined whether cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) the markers nephrin, podocin, SIX2, OSR1, LHX1, RET, FGF8, and RACK1.

[0024] In a method of identifying an enriched heterogeneous renal cell population having therapeutic potential, determining that cells of the heterogeneous enriched renal cell population express (i) NEPH1 and (ii) at least one (e.g., any one, or any two, or any three, or any four, or any five, or any six, or any seven, or all eight) of the markers can identify the enriched heterogeneous renal cell population as having therapeutic potential.

[0025] In a method of identifying an enriched heterogeneous renal cell population having therapeutic potential, determining the expression of (i) NEPH1 and (ii) one or more (or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers may further comprise determining the percentage of cells of the enriched heterogeneous renal cell population that express (i) NEPH1 and / or (ii) one or more (or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers. If the percentage of cells of the enriched heterogeneous renal cell population that express (i) NEPH1 and / or (ii) one or more (or at least one, at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, then the enriched heterogeneous renal cell population can be identified as having therapeutic potential if a certain or specific percentage of the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and / or (ii) one or more (or at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers.

[0026] In the method, if the percentage of cells of the enriched heterogeneous renal cell population that express (i) NEPH1 is determined, then the enriched heterogeneous renal cell population can be identified as having therapeutic potential if about 2.0% to about 8.0%, or about 2.0% to about 7.5%, or about 2.0% to about 7.0%, or about 2.0% to about 6.5%, or about 2.0% to about 6.0%, or about 2.0% to about 5.5%, or about 2.0% to about 5.0%, or about 2.5% to about 8.0%, or about 3.0% to about 8.0%, or about 3.5% to about 8.0%, or about 4.0% to about 8.0%, or about 2.5% to about 7.5%, or about 3.0% to about 7.0%, or about 3.5% to about 6.5%, or about 4.0% to about 6.0%, or about 4.5% to about 5.5%, or about 4.0% to about 6.5%, or about 4.0% to about 7.0%, or about 3.5% to about 6.5%, or about 3.5% to about 6.0%, or about 3% to about 6.0% of the cells of the enriched heterogeneous population express NEPH1.

[0027] In this method, if the percentage of cells in an enriched heterogeneous renal cell population expressing (ii) one or more (or at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include SIX2, then if at least about 10% of the cells in the enriched heterogeneous renal cell population express SIX2, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Alternatively, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include SIX2, then if at least about 0.15%, or at least about 0.20%, or at least about 0.50%, or at least about 1.0%, or at least about 1.5%, or at least about 2.0%, or at least about 2.5%, or at least about 3.0%, or at least about 3.5%, or at least about 4.0%, or at least about 4.5%, or at least about 5.0%, or at least about 5.5% of the cells in the enriched heterogeneous renal cell population express SIX2, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. If the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include SIX2, then if more than 0% and up to about 35.0%, more than 0% and up to about 30.0%, or more than 0% and up to about 25.0%, or more than 0% and up to about 20.0%, or more than 0% and up to about 15.0%, or more than 0% and up to about 12.5%, or more than 0% and up to about 10.0%, or more than 0% and up to about 7.5%, or more than 0% and up to about 5.0% of the cells in the enriched heterogeneous renal cell population express SIX2, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.In addition, if one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include SIX2, then if about 0.10% to about 15.0%, or about 0.10% to about 10.0%, or about 0.10% to about 8.0%, or about 0.10% to about 6.0%, or about 0.10% to about 4.0%, or about 0.10% to about 2.0%, or about 0.40% to about 15.0%, or about 0.40% to about 10.0%, or about 0.40% to about 8.0%, or about 0.40% to about 6.0%, or about 0.40% to about 4.0%, or about 0.40% to about 2.0%, or about 0.60% to about 15.0%, or about 0.60% to about 10.0%, or about 0.60% to about 8.0%, or about 0.60% to about 6.0%, or about 0.60% to about 4.0%, or about 0.60% to about 2.0% of the cells in the enriched heterogeneous renal cell population express SIX2, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0028] In this method, if the percentage of cells in an enriched heterogeneous renal cell population that express one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include OSR1, then if at least about 35% of the cells in the enriched heterogeneous renal cell population express OSR1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Alternatively, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include OSR1, then if at least about 45%, or at least about 47.5%, or at least about 50%, or at least about 52.5%, or at least about 55%, or at least about 57.5%, or at least about 60%, or at least about 62.5%, or at least about 65%, or at least about 67.5%, or at least about 70%, or at least about 72.5%, or at least about 75% of the cells in the enriched heterogeneous renal cell population express OSR1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. If the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include OSR1, then if about 35% to about 99%, or about 35% to about 98%, or about 35% to about 96%, or about 35% to about 94%, or about 35% to about 92%, or about 35% to about 90%, or about 40% to 99%, or about 40% to about 98%, or about 40% to about 96%, or about 40% to about 94%, or about 40% to about 92%, or about 40% to about 90%, or about 45% to about 99%, or about 45% to about 98%, or about 45% to about 96%, or about 45% to about 94%, or about 45% to about 92%, or about 45% to about 90%, or about 50% to about 99%, or about 50% to about 98%, or about 50% to about 96%, or about 50% to about 94%, or about 50% to about 92%, or about 50% to about 90%, or about 55% to about 99%, or about 55% to about 98%, or about 55% to about 96%, or about 55% to about 94%, or about 55% to about 92%, or about 55% to about 90%, or about 60% to about 99%, or about 60% to about 98%, or about 60% to about 96%, or about 60% to about 94%, or about 60% to about 92%, or about 60% to about 90% of the cells in the enriched heterogeneous renal cell population express OSR1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0029] In this method, if the percentage of cells in an enriched heterogeneous renal cell population that express one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include LHX1, then if at least about 0.80% of the cells in the enriched heterogeneous renal cell population express LHX1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Alternatively, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include LHX1, then if at least about 1%, or at least about 3%, or at least about 5%, or at least about 7%, or at least about 9%, or at least about 11%, or at least about 13%, or at least about 15% of the cells in the enriched heterogeneous renal cell population express LHX1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. If the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include LHX1, then if greater than 0% and up to 99%, or greater than 0% and up to 90%, or greater than 80% and up to 75%, or greater than 0% and up to about 70%, or greater than 0% and up to about 65%, or greater than 0% and up to about 60%, or greater than 0% and up to about 55%, or greater than 0% and up to about 50%, or greater than 0% and up to about 45%, or greater than 0% and up to about 40%, or greater than 0% and up to about 35%, or greater than 0% and up to about 30% of the cells in the enriched heterogeneous renal cell population express LHX1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.In addition, if one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include LHX1, then if about 0.80% to about 99%, or about 0.80% to about 90%, or about 0.80% to about 80%, or about 0.80% to about 70%, or about 0.80% to about 60%, or about 0.80% to about 50%, or about 0.80% to about 40%, or about 5% to about 99%, or about 5% to about 90%, or about 5% to about 80%, or about 5% to about 70%, or about 5% to about 60%, or about 5% to about 50%, or about 5% to about 40%, or about 10% to about 99%, or about 10% to about 90%, or about 10% to about 80%, or about 10% to about 70%, or about 10% to about 60%, or about 10% to about 50%, or about 10% to about 40% of the cells in the enriched heterogeneous renal cell population express LHX1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0030] In this method, if the percentage of cells in an enriched heterogeneous renal cell population expressing one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include RET, then if at least about 1% of the cells in the enriched heterogeneous renal cell population express RET, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Alternatively, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include RET, then if at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11% or at least about 12% of the cells in the enriched heterogeneous renal cell population express RET, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. If the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include RET, then if more than 1% and up to about 85%, or more than 1% and up to about 83%, or more than 1% and up to about 80%, or more than 1% and up to about 77%, or more than 1% and up to about 75%, or more than 1% and up to about 72%, or more than 1% and up to about 70%, or more than 1% and up to about 67%, or more than 1% and up to about 65%, or more than 1% and up to about 63%, or more than 1% and up to about 60% of the cells in the enriched heterogeneous renal cell population express RET, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Further, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include RET, then if about 1% to about 80%, or about 1% to about 77%, or about 2% to about 80%, or about 2% to about 77%, or about 4% to about 80%, or about 4% to about 78%, or about 5% to about 70%, or about 5% to about 65%, or about 5% to about 60%, or about 10% to about 80%, or about 10% to about 75%, or about 10% to about 70%, or about 10% to about 65% or about 10% to about 60% of the cells in the enriched heterogeneous renal cell population express RET, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0031] In this method, if the percentage of cells in an enriched heterogeneous renal cell population that express one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include FGF8, then if at least about 0.01% of the cells in the enriched heterogeneous renal cell population express FGF8, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Alternatively, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include FGF8, then if at least about 0.01%, or at least about 0.03%, or at least about 0.05%, or at least about 0.07%, or at least about 0.09%, or at least about 0.10%, or at least about 0.20%, or at least about 0.30%, or at least about 0.40%, or at least about 0.50%, or at least about 0.60%, or at least about 0.70%, or at least about 0.80% or at least about 0.90% of the cells in the enriched heterogeneous renal cell population express FGF8, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. If the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include FGF8, then if greater than 0% and up to about 60%, or greater than 0% and up to about 57%, or greater than 0% and up to about 55%, or greater than 0% and up to about 52%, or greater than 0% and up to about 50%, or greater than 0% and up to about 47%, or greater than 0% and up to about 45%, or greater than 0% and up to about 42%, or greater than 0% and up to about 40%, or greater than 0% and up to about 37% or greater than 0% and up to about 35% of the cells in the enriched heterogeneous renal cell population express FGF8, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.In addition, if one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include FGF8, then if from about 0.01% to about 60%, or from about 0.01% to about 58%, or from about 0.01% to about 50%, or from about 0.01% to about 45%, or from about 0.01% to about 40%, or from about 0.01% to about 35%, or from about 0.01% to about 30%, or from about 0.01% to about 25%, 0.10% to about 60%, or from about 0.10% to about 58%, or from about 0.10% to about 50%, or from about 0.10% to about 45%, or from about 0.10% to about 40%, or from about 0.10% to about 35%, or from about 0.10% to about 30%, or from about 0.10% to about 25%, 0.20% to about 60%, or from about 0.20% to about 58%, or from about 0.20% to about 50%, or from about 0.20% to about 45%, or from about 0.20% to about 40%, or from about 0.20% to about 35%, or from about 0.20% to about 30%, or from about 0.20% to about 25% of the cells in the enriched heterogeneous renal cell population express FGF8, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0032] In this method, if the percentage of cells in an enriched heterogeneous renal cell population that express one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include RACK1, then if at least about 75% of the cells in the enriched heterogeneous renal cell population express RACK1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Alternatively, if one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include RACK1, then if at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% of the cells in the enriched heterogeneous renal cell population express RACK1, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0033] In this method, if the percentage of cells in an enriched heterogeneous renal cell population that express one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include nephropin, then if at least about 65% of the cells in the enriched heterogeneous renal cell population express nephropin, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. Alternatively, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include nephropin, then if at least about 68%, at least about 70%, at least about 72%, at least about 74%, at least about 76%, at least about 78%, at least about 80%, at least about 82%, at least about 84%, at least about 86%, at least about 88%, at least about 90%, at least about 92%, at least about 94%, at least about 96%, at least about 98% or about 100% of the cells in the enriched heterogeneous renal cell population express nephropin, the enriched heterogeneous renal cell population can be identified as having therapeutic potential. If the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include nephropin, then if about 65% to about 99%, or about 67% to about 99%, or about 69% to about 99%, or about 70% to about 99%, or about 72% to about 99%, about 74% to about 99%, or 35% to about 95%, or about 40% to about 95%, or about 45% to 95%, or about 50% to about 95%, or about 55% to about 95%, or about 60% to about 95%, or about 65% to about 95%, or 70% to about 95%, or about 75% to about 95%, or about 80% to about 95% or about 85% to about 95% of the cells in the enriched heterogeneous renal cell population express nephropin, the enriched heterogeneous renal cell population can be identified as having therapeutic potential.

[0034] In this method, if the percentage of cells in an enriched heterogeneous renal cell population that express one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, and the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include podocin, then the enriched heterogeneous renal cell population can be identified as having therapeutic potential if at least about 90% of the cells in the enriched heterogeneous renal cell population express podocin. Alternatively, if the one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers are or include podocin, then the enriched heterogeneous renal cell population can be identified as having therapeutic potential if at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 98%, at least about 98% or at least about 100% of the cells in the enriched heterogeneous renal cell population express podocin.

[0035] In this method, if the percentage of cells in a heterogeneous renal cell population expressing (i) NEPH1 and (ii) one or more (at least one, or at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or eight) markers is determined, then the percentage of cells expressing (i) NEPH1 and (ii) any two, any three, any four, any five, any six, any seven, or all eight of the markers SIX2, OSR1, LHX1, RET, FGF8, RACK1, nephroprotein, and podocin in any combination can identify it as having therapeutic potential. For example, if (a) it is determined that about 2% to about 8.0%, about 3.0% to about 7.0%, or about 4.0% to about 6.0% of the cells in a heterogeneous renal cell population express NEPH1 and (b) any combination of any two, any three, any four, any five, any six, any seven, or all eight of the following is determined: (i) greater than 0% and up to about 35.0% of the cells in the heterogeneous renal cell population express SIX2, (ii) at least about 35% of the cells in the heterogeneous renal cell population express OSR1, (iii) at least about 1% of the cells in the heterogeneous renal cell population express LHX1, (iv) at least about 1% of the cells in the heterogeneous renal cell population express RET, (v) greater than 0% and up to about 60% of the cells in the heterogeneous renal cell population express FGF8, (vi) at least about 80% of the cells in the heterogeneous renal cell population express RACK1, (vii) at least about 65% of the cells in the enriched heterogeneous renal cell population express nephroprotein, and / or (viii) about 90% of the cells in the enriched heterogeneous renal cell population express podocin. In another example, if (a) it is determined that about 2% to about 8.0%, about 3.0% to about 7.0%, or about 4.0% to about 6.0% of the cells in a heterogeneous renal cell population express NEPH1 and (b) any combination of any two, any three, any four, any five, any six,Any combination of seven or all eight allows the enriched heterogeneous renal cell population to be identified as having therapeutic potential: (i) from about 0.1% to about 30.0% of the cells in the heterogeneous renal cell population express SIX2, (ii) from about 40% to about 99% of the cells in the heterogeneous renal cell population express OSR1, (iii) at least about 1% of the cells in the heterogeneous renal cell population express LHX1, (iv) from about 1% to about 80% of the cells in the heterogeneous renal cell population express RET, (v) from about 0.01% to about 60% of the cells in the heterogeneous renal cell population express FGF8, (vi) at least about 80% of the cells in the enriched heterogeneous renal cell population express RACK1, (vii) at least about 65% of the cells in the enriched heterogeneous renal cell population express nephrin, and / or (viii) about 90% of the cells in the enriched heterogeneous renal cell population express podocin. The expression can be determined as a combination of any two, any three, any four, any five, any six, any seven, or all eight of these percentages and can be any of the following combinations: SIX2 and OSR1; SIX2 and LHX1; SIX2 and RET; SIX2 and FGF8; OSR1 and LHX1; OSR1 and RET; OSR1 and FGF8; LHX1 and RET; LHX1 and FGF8; RET and FGF8; nephrin and SIX2; nephrin and OSR1; nephrin and LHX1, nephrin and RET, nephrin and FGF8; podocin and nephrin; podocin and SIX2; podocin and OSR1; podocin and LHX1; podocin and RET; podocin and FGF8; RACK1 and nephrin; RACK1 and podocin; RACK1 and SIX2; RACK1 and OSR1; RACK1 and LHX1; RACK1 and RET; RACK1 and FGF8; SIX2, OSR1 and LHX1; SIX2, OSR1 and RET; SIX2, OSR1 and FGF8; SIX2, LHX1 and RET; SIX2, LHX1 and FGF8; SIX2, RET and FGF8; OSR1, LHX1 and RET; OSR1, LHX1 and FGF8; OSR1, RET and FGF8; LHX1, RET and FGF8; nephrin, SIX2 and OSR; nephrin, SIX2 and LHX1; nephrin, SIX2 and RET; nephrin, SIX2 and FGF8; nephrin, OSR and LHX1; nephrin, OSR and RET; nephrin, OSR and FGF8; nephrin, LHX1 and RET; nephrin, LHX1 and FGF8; nephrin, RET and FGF8; podocin, SIX2 and OSR; podocin, SIX2 and LHX1; podocin, SIX2 and RET; podocin, SIX2 and FGF8; podocin, OSR and LHX1; podocin, OSR and RET; podocin,OSR and FGF8; Nephrin, LHX1 and RET; Nephrin, LHX1 and FGF8; Nephrin, RET and FGF8; RACK1, SIX2 and OSR; RACK1, SIX2 and LHX1; RACK1, SIX2 and RET; RACK1, SIX2 and FGF8; RACK1, OSR and LHX1; RACK1, OSR and RET; RACK1, OSR and FGF8; RACK1, LHX1 and RET; RACK1, LHX1 and FGF8; RACK1, RET and FGF8; Nephrin, Nephrin and RACK1; Nephrin, Nephrin and SIX2; Nephrin, Nephrin and OSR; Nephrin, Nephrin and RET; Nephrin, Nephrin and LHX1; Nephrin, Nephrin and FGF8; Nephrin, RACK1 and SIX2; Nephrin, RACK1 and OSR; Nephrin, RACK1 and RET; Nephrin, RACK1 and LHX1; Nephrin, RACK1 and FGF8; Nephrin, RACK1 and SIX2; Nephrin, RACK1 and OSR; Nephrin, RACK1 and RET; Nephrin, RACK1 and LHX1; Nephrin, RACK1 and FGF8; SIX2, OSR1, LHX1 and RET; SIX2, OSR1, LHX1 and FGF8; SIX2, LHX1, RET and FGF8; SIX2, OSR1, RET and FGF8; OSR1, LHX1, RET and FGF8; Nephrin, SIX2, OSR1 and LHX1; Nephrin, SIX2, OSR1 and RET; Nephrin, SIX2, OSR1 and FGF8; Nephrin, SIX2, LHX1 and RET; Nephrin, SIX2, LHX1 and FGF8; Nephrin, SIX2, RET and FGF8; Nephrin, OSR1, LHX1 and FGF8; Nephrin, OSR1, LHX1 and RET; Nephrin, OSR1, RET and FGF8; Nephrin, LHX1, RET and FGF8; Nephrin, SIX2, OSR1 and LHX1; Nephrin, SIX2, OSR1 and RET; Nephrin, SIX2, OSR1 and FGF8; Nephrin, SIX2, LHX1 and RET; Nephrin, SIX2, LHX1 and FGF8; Nephrin, SIX2, RET and FGF8; Nephrin, OSR1, LHX1 and FGF8; Nephrin, OSR1, LHX1 and RET; Nephrin, OSR1, RET and FGF8; Nephrin, LHX1, RET and FGF8; RACK1, SIX2, OSR1 and FGF8; RACK1, SIX2, LHX1 and RET; RACK1, SIX2, LHX1 and FGF8; RACK1, SIX2, RET and FGF8; RACK1,OSR1, LHX1, and FGF8; RACK1, OSR1, LHX1, and RET; RACK1, OSR1, RET, and FGF8; RACK1, LHX1, RET, and FGF8; Nephrin, Podocin, SIX2, and OSR1; Nephrin, Podocin, SIX2, and LHX1; Nephrin, Podocin, SIX2, and RET; Nephrin, Podocin, SIX2, and FGF8; Nephrin, Podocin, OSR1, and LHX1; Nephrin, Podocin, OSR1, and RET; Nephrin, Podocin, OSR1, and FGF8; Nephrin, Podocin, LHX1, and RET; Nephrin, Podocin, LHX1, and FGF8; Nephrin, Podocin, RET, and FGF8; RACK1, Podocin, SIX2, and OSR1; RACK1, Podocin, SIX2, and LHX1; RACK1, Podocin, SIX2, and RET; RACK1, Podocin, SIX2, and FGF8; RACK1, Podocin, OSR1, and LHX1; RACK1, Podocin, OSR1, and RET; RACK1, Podocin, OSR1, and FGF8; RACK1, Podocin, LHX1, and RET; RACK1, Podocin, LHX1, and FGF8; RACK1, Podocin, RET, and FGF8; RACK1, Nephrin, SIX2, and OSR1; RACK1, Nephrin, SIX2, and LHX1; RACK1, Nephrin, SIX2, and RET; RACK1, Nephrin, SIX2, and FGF8; RACK1, Nephrin, OSR1, and LHX1; RACK1, Nephrin, OSR1, and RET; RACK1, Nephrin, OSR1, and FGF8; RACK1, Nephrin, LHX1, and RET; RACK1, Nephrin, LHX1, and FGF8; RACK1, Nephrin, RET, and FGF8; RACK1, Nephrin, Podocin, and SIX2; RACK1, Nephrin, Podocin, and RET; RACK1, Nephrin, Podocin, and LHX1; RACK, Nephrin, Podocin, and OSR1; RACK, Nephrin, Podocin, and FGF8; SIX2, OSR1, LHX1, RET, and FGF8; Nephrin, SIX2, OSR1, LHX1, and RET; Nephrin, SIX2, OSR1, LHX1, and FGF8; Nephrin, SIX2, LHX1, RET, and FGF8; Nephrin, SIX2, OSR1, RET, and FGF8; Nephrin, OSR1, LHX1, RET, and FGF8; Podocin, SIX2, OSR1, LHX1, and RET; Podocin, SIX2, OSR1, LHX1, and FGF8; Podocin, SIX2, LHX1, RET, and FGF8; Podocin, SIX2, OSR1,RET and FGF8; podocin, OSR1, LHX1, RET and FGF8; RACK1, SIX2, OSR1, LHX1 and RET; RACK1, SIX2, OSR1, LHX1 and FGF8; RACK1, SIX2, LHX1, RET and FGF8; RACK1, SIX2, OSR1, RET and FGF8; RACK1, OSR1, LHX1, RET and FGF8; nephrin, podocin, SIX2, OSR1 and LHX1; nephrin, podocin, SIX2, OSR1 and RET; nephrin, podocin, SIX2, OSR1 and FGF8; nephrin, podocin, SIX2, LHX1 and RET; nephrin, podocin, SIX2, LHX1 and FGF8; nephrin, podocin, SIX2, RET and FGF8; nephrin, podocin, OSR1, LHX1 and RET; nephrin, podocin, OSR1, LHX1 and FGF8; nephrin, podocin, OSR1, RET and FGF8; nephrin, podocin, LHX1, RET and FGF8; nephrin, RACK1, SIX2, OSR1 and LHX1; nephrin, RACK1, SIX2, OSR1 and RET; nephrin, RACK1, SIX2, OSR1 and FGF8; nephrin, RACK1, SIX2, LHX1 and RET; nephrin, RACK1, SIX2, LHX1 and FGF8; nephrin, RACK1, SIX2, RET and FGF8; nephrin, RACK1, OSR1, LHX1 and RET; nephrin, RACK1, OSR1, LHX1 and FGF8; nephrin, RACK1, OSR1, RET and FGF8; nephrin, RACK1, LHX1, RET and FGF8; podocin, RACK1, SIX2, OSR1 and LHX1; podocin, RACK1, SIX2, OSR1 and RET; podocin, RACK1, SIX2, OSR1 and FGF8; podocin, RACK1, SIX2, LHX1 and RET; podocin, RACK1, SIX2, LHX1 and FGF8; podocin, RACK1, SIX2, RET and FGF8; podocin, RACK1, OSR1, LHX1 and RET; podocin, RACK1, OSR1, LHX1 and FGF8; podocin, RACK1, OSR1, RET and FGF8; podocin, RACK1, LHX1, RET and FGF8; nephrin, podocin, RACK1, SIX2 and OSR1; nephrin, podocin, RACK1, SIX2 and LHX1; nephrin, podocin, RACK1, SIX2 and RET; nephrin, podocin, RACK1, SIX2 and FGF8; nephrin, podocin,RACK1, OSR1, and LHX1; Nephrin, Podocin, RACK1, OSR1, and RET; Nephrin, Podocin, RACK1, OSR1, and FGF8; Nephrin, Podocin, RACK1, LHX1, and RET; Nephrin, Podocin, RACK1, LHX1, and FGF8; Nephrin, Podocin, RACK1, RET, and FGF8; Nephrin, SIX2, OSR1, LHX1, RET, and FGF8; Podocin, SIX2, OSR1, LHX1, RET, and FGF8; RACK1, SIX2, OSR1, LHX1, RET, and FGF8; Nephrin, Podocin, SIX2, OSR1, LHX1, and RET; Nephrin, Podocin, SIX2, OSR1, LHX1, and FGF8; Nephrin, Podocin, SIX2, LHX1, RET, and FGF8; Nephrin, Podocin, SIX2, OSR1, RET, and FGF8; Nephrin, Podocin, OSR1, LHX1, RET, and FGF8; Nephrin, RACK1, SIX2, OSR1, LHX1, and RET; Nephrin, RACK1, SIX2, OSR1, LHX1, and FGF8; Nephrin, RACK1, SIX2, LHX1, RET, and FGF8; Nephrin, RACK1, SIX2, OSR1, RET, and FGF8; Nephrin, RACK1, OSR1, LHX1, RET, and FGF8; Podocin, RACK1, SIX2, OSR1, LHX1, and RET; Podocin, RACK1, SIX2, OSR1, LHX1, and FGF8; Podocin, RACK1, SIX2, LHX1, RET, and FGF8; Podocin, RACK1, SIX2, OSR1, RET, and FGF8; Podocin, RACK1, OSR1, LHX1, RET, and FGF8; Nephrin, Podocin, RACK1, SIX2, OSR1, and LHX1; Nephrin, Podocin, RACK1, SIX2, OSR1, and RET; Nephrin, Podocin, RACK1, SIX2, OSR1, and FGF8; Nephrin, Podocin, RACK1, SIX2, LHX1, and RET; Nephrin, Podocin, RACK1, SIX2, LHX1, and FGF8; Nephrin, Podocin, RACK1, SIX2, RET, and FGF8; Nephrin, Podocin, RACK1, OSR1, LHX1, and RET; Nephrin, Podocin, RACK1, OSR1, LHX1, and FGF8; Nephrin, Podocin, RACK1, OSR1, RET, and FGF8; Nephrin, Podocin, RACK1, LHX1, RET, and FGF8; Podocin, RACK1, SIX2, OSR1, LHX1, RET, and FGF8; Nephrin,RACK1, SIX2, OSR1, LHX1, RET, and FGF8; nephrin, podocin, SIX2, OSR1, LHX1, RET, and FGF8; nephrin, podocin, RACK1, OSR1, LHX1, RET, and FGF8; nephrin, podocin, RACK1, SIX2, LHX1, RET, and FGF8; nephrin, podocin, RACK1, SIX2, OSR1, RET, and FGF8; nephrin, podocin, RACK1, SIX2, OSR1, LHX1, and FGF8; nephrin, podocin, RACK1, SIX2, OSR1, LHX1, and RET; or SIX2, OSR1, LHX1, RET, FGF8, RACK1, nephrin, and podocin.

[0036] In a method of identifying an enriched heterogeneous renal cell population having therapeutic potential, it can be determined whether the cells of the enriched heterogeneous renal cell population express one or more further markers, i.e., markers other than NEPH1, nephrin, podocin, RACK1, SIX2, OSR1, LHX1, RET, and / or FGF8. One or more further markers can be or can include one or more of lefty, nodal, or CD24.

[0037] It should be understood that if the percentage of cells expressing a certain marker is expressed as a percentage “about” a particular number (e.g., about 5%), the percentage of cells does not have to be the exact particular number, e.g., exactly 5%. Instead, it should be understood that if the percentage of cells expressing a certain marker is expressed as a percentage “about” a particular number, e.g., about 5%, the percentage of cells expressing the particular marker can be within a range of up to 10% of that particular number, e.g., between 4.5% and 5.5%.

[0038] In any method for identifying an enriched heterogeneous renal cell population having therapeutic potential, determining whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) one or more markers (e.g., SIX2, OSR1, LHX1, FGF8, RACK1, RET, nephrocystin, and / or podocin) and optionally (iii) one or more further markers can involve determining whether the cells of the enriched heterogeneous renal cell population express (i) NEPH1 and (ii) one or more markers and optionally (iii) one or more further markers in the form of nucleic acids (e.g., mRNA or miRNA) or polypeptides. Expression of (i) NEPH1 and (ii) one or more markers (e.g., SIX2, OSR1, LHX1, FGF8, RACK1, RET, nephrocystin, and / or podocin) and optionally (iii) one or more further markers in the cells of the enriched heterogeneous renal cell population can be determined via any assay suitable for detecting the presence of (i) NEPH1 and (ii) one or more markers and optionally (iii) one or more further markers. For example, if expression is determined in polypeptide form, it can be determined by assays such as Western blotting, fluorescence-activated cell sorting (FACS), enzyme-linked immunosorbent assay (ELISA). If expression is determined in nucleic acid form, it can be determined by assays such as or using Southern blotting, polymerase chain reaction (PCR) or reverse transcriptase PCR, serial analysis of gene expression (SAGE), MassARRAY, or fluorescence in situ hybridization (FISH). The assay can be an assay using labeled detection reagents to determine the presence of (i) NEPH1 and (ii) one or more markers and optionally (iii) one or more further markers and / or the percentage of cells expressing (i) NEPH1 and (ii) one or more markers and optionally (iii) one or more further markers. The labeled detection reagents can include (a) a moiety directly or indirectly conjugated to NEPH1 and one or more markers and / or further markers and (b) a detection moiety. Non-limiting detection moieties include radioisotopes, e.g., 35 S, 14 C, 125 I, 3 H, and 131 I, colloidal gold particles, fluorescent labels, e.g., Texas Red, rhodamine, fluorescein, dansyl, Lissamine, phycoerythrin, phycocyanin, SPECTRUM ORANGE, SPECTRUM GREEN1, and enzyme substrates, e.g., firefly luciferase, bacterial luciferase, luciferin, horseradish peroxidase, alkaline phosphatase, or β-galactosidase.

[0039] An enriched heterogeneous renal cell population that can be identified as having therapeutic potential in any method can enrich one or more renal cell types, such as renal epithelial cells, renal tubular cells, renal tubular epithelial cells, or renal proximal tubular cells. The enrichment of the enriched heterogeneous renal cell population for these one or more renal cell types can mean that the enriched heterogeneous renal cell population has a greater percentage of one or more renal cell types than the patient's renal tissue, the patient's renal biopsy, or an in vitro culture of cells established from the patient's renal tissue or renal biopsy (which can be collectively referred to as the "starting renal cell population"). The starting renal cell population, if it is an in vitro culture of cells established from the patient's renal tissue or the patient's renal biopsy, can be a renal cell preparation containing dissociated cells of the renal tissue or renal biopsy (e.g., cells dissociated from the renal tissue or renal biopsy via mincing and / or enzymatic digestion), which may have been processed to remove red blood cells and debris or may not have been processed. In addition to enriching renal epithelial cells, renal tubular cells, renal tubular epithelial cells, and / or renal proximal tubular cells, the enriched heterogeneous renal cell population can also include other renal cell types, such as glomerular cells, podocytes, collecting duct cells, and / or vascular cells.

[0040] An enriched heterogeneous renal cell population can be prepared from a starting renal cell population (e.g., 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) by a method that includes a separation step to enrich one or more renal cell types. The separation step can be a separation step of separating the cells of the starting renal cell population that have been passaged no more than once, twice, or three times based on their buoyant density. If the separation step is to separate cells based on the buoyant density of the cells, the separation step can utilize a single-step or multi-step continuous or discontinuous density gradient using a density gradient medium such as glycerol, OptiPrep, Percoll, or Ficoll-Paque. Using such density gradient media in this way can result in the separation of the cells of the starting renal cell population (or the starting renal cell population that has been passaged at most once, twice, or three times) into one or more distinguishable fractions from which the cells of the enriched heterogeneous renal cell population can be clearly identified and separated. The distinguishable fraction(s) can be the fraction(s) in which the buoyant density of the cells in the fraction 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. The distinguishable fraction(s) can be the fraction(s) in which the buoyant density of the cells in the fraction 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. The distinguishable fraction(s) can be the fraction(s) 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 of separating the cells of the starting renal cell population (or the cells of the starting renal cell population that has been passaged no more than once, twice, or three times) based on whether they express a specific marker on their surface. If the separation step is to separate cells based on the expression of their specific cell surface markers, the separation step can be a separation step using flow cytometry. If the cells express a specific surface marker such as nephrin (e.g., characteristic of renal epithelial cells, renal tubular cells, renal tubular epithelial cells, or renal proximal tubular cells), thereby forming a separated enriched heterogeneous renal cell population, flow cytometry can sort the cells from the starting renal cell population (or the starting renal cell population that has been passaged at most once, twice, or three times).

[0041] The enriched heterogeneous renal cell population that has been prepared from the starting renal cell population (or the starting renal cell population that has been passaged at most once, twice, or three times) can be cultured under hypoxic conditions prior to the separation step. If the cells are cultured under hypoxic conditions prior to the separation step, the cells 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% oxygen, or less than about 4% oxygen, or less than about 3% oxygen, or less than about 2% oxygen. If the 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, or at least 48 hours.

[0042] Generally, the preparation of the enriched heterogeneous renal cell population can be from any starting cell population, for example, an in vitro culture of cells established from a patient's renal 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 renal tissue or renal biopsy, the cells of the in vitro culture can be expanded by passaging at most once, or at most twice, or at most three times. Alternatively, if desired, the cells of the in vitro culture established from renal tissue or renal biopsy can be cryopreserved and then expanded by passaging at most once, or at most twice, or at most three times. Once the cells are expanded, the expanded cells can be cryopreserved. The expanded cells, whether cryopreserved or not, can then undergo the separation step or can then undergo hypoxic culture conditions followed by the separation step. The enriched heterogeneous renal cell population can be separated by performing the separation step. Once the enriched heterogeneous renal cell population is isolated, it can be frozen and / or analyzed prior to being used as a therapeutic agent.

[0043] If the enriched heterogeneous renal cell population is identified as having therapeutic potential according to any of the methods disclosed herein, it can be included in a pharmaceutical composition or administered in a method of treating a kidney disease in a patient in need thereof, and / or used in the preparation of a medicament for treating a kidney disease. If the enriched heterogeneous renal cell population is identified as having therapeutic potential and is included in a pharmaceutical composition, it can be formulated as a hydrogel composition or a liquid composition. The pharmaceutical composition form can contain or not contain hyaluronic acid.

[0044] If the pharmaceutical composition is formulated as a hydrogel composition, the cells of the enriched heterogeneous renal cell composition can be combined with a temperature-sensitive cell-stabilizing biomaterial. The temperature-sensitive cell-stabilizing biomaterial can be a biomaterial that is in a gel state at certain temperatures and in a liquid state at other temperatures. For example, if the biomaterial is temperature-sensitive, the biomaterial can be in a gel state at about 8°C or below, substantially in a liquid state at about room temperature or above, and in a solid-liquid transition state between about 8°C and room temperature; or in a gel state at about 4°C or below, in a liquid state at about 37°C or above, and in a solid-liquid transition state between about 8°C and about 18°C; or in a gel state at about 2°C or below, in a liquid state at about 37°C or above, and in a solid-liquid transition state between about 8°C and about 18°C; or in a gel state at about 2°C or below and in a liquid state at about 37°C or above; or in a gel state at about 4°C or below and in a liquid state at about 34°C or above; or in a gel state at about 6°C or below and in a liquid state at about 32°C or above. The temperature-sensitive cell-stabilizing biomaterial can include, or consist of, one or more proteins or peptides of natural origin or recombinant origin. The protein or peptide of natural origin or recombinant origin can be an extracellular matrix protein of recombinant origin, or an extracellular matrix derived from the kidney or another tissue or organ, or gelatin. If the temperature-sensitive cell-stabilizing biomaterial is or contains gelatin, the gelatin can 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 contains gelatin, the gelatin can be present in the therapeutic composition at about 0.5% to about 1% 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 cells of the enriched heterogeneous renal cell population can be dispersed throughout the biomaterial, or be distributed substantially uniformly throughout the biomaterial. The cells of the enriched heterogeneous renal cell population can be formulated in a biomaterial such as gelatin such that the number of cells per mL of biomaterial is about 20 x 10 6 cells per mL, about 40 x 10 6 cells per mL, about 60 x 10 6 cells per mL, about 100 x 10 6 cells per mL, about 120 x10 6 cells per mL, about 140 x 10 6 cells per mL, about 160 x 10 6 cells per mL, about 180 x 10 6 cells per mL, or about 200 x 10 6 cells per mL.

[0045] 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 cryopreservation until the time of 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, glucose, 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 the liquid is about 20 x 10 6 cells per mL, about 40 x 10 6 cells per mL, about 60 x 10 6 cells per mL, about 100 x 10 6 cells per mL, about 120 x 10 6 cells per mL, about 140 x 10 6 cells per mL, about 160 x 10 6 cells per mL, about 180 x 10 6 cells per mL, or about 200 x 10 6 cells per mL.

[0046] If the enriched heterogeneous renal cell population is identified as having therapeutic potential, the enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can be administered to a patient in a method of treating a kidney disease or can be used in a method of treating a kidney disease. If the enriched heterogeneous renal cell population is identified as having therapeutic potential, the enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can be used to prepare a medicament for treating a kidney disease. The kidney disease can be any stage or degree of acute or chronic renal failure. It may originate in the kidney or it may be secondary to another condition, such as heart failure, hypertension, diabetes, an autoimmune disease, or a liver disease. Alternatively, the kidney disease can be a kidney disease caused by an acute injury to the kidney or a result of an abnormality of the kidney and / or urinary tract. The kidney disease can further include endocrine dysfunctions such as anemia (e.g., erythropoietin deficiency) and mineral imbalances (e.g., vitamin D deficiency).

[0047] If an enriched heterogeneous renal cell population is identified as having therapeutic potential, then administering the enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can treat kidney diseases. It can treat kidney diseases by restoring renal function, stabilizing renal function, improving renal function, reducing renal fibrosis, or reducing renal inflammation in the kidneys of patients in need of such treatment. Treating kidney diseases can restore mineral balance or relieve anemia in patients in need of such treatment. Treating kidney diseases can delay or avoid the need for dialysis, or can delay or avoid the need for kidney transplantation in patients in need of treatment for kidney diseases. If treating kidney diseases delays a patient's need for dialysis or kidney transplantation, the delay can be at least 1 year, at least 1.5 years, at least 2 years, at least 2.5 years, at least 3 years, at least 3.5 years, at least 4 years, at least 4.5 years, at least 5 years, at least 5.5 years, at least 6 years, at least 6.5 years, at least 7 years, at least 7.5 years, at least 8 years, at least 8.5 years, at least 9 years, at least 9.5 years, or at least 10 years. Treatment of kidney diseases can be determined by observing improvements in a patient's serum albumin, albumin to globulin ratio (A / G ratio), serum phosphorus, serum sodium, kidney size (measured by ultrasound), serum calcium, phosphorus:calcium ratio, serum potassium, proteinuria, urinary creatinine, serum creatinine, blood urea nitrogen (BUN), cholesterol level, triglyceride level, and glomerular filtration rate (GFR), body weight, blood pressure (mean systemic blood pressure, diastolic blood pressure, or systolic blood pressure), and physical endurance performance.

[0048] If an enriched heterogeneous renal cell population is identified as having therapeutic potential, it can be administered to a patient by any suitable route of administration known in the art. For example, the enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can be administered systemically to a patient in need of treatment for kidney diseases. The enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can be administered at or into the kidney of a patient in need of treatment for kidney diseases. If the enriched heterogeneous renal cell population is administered at or into the kidney of a patient in need of treatment for kidney diseases, it can be administered by single or multiple injections. It can be administered via direct laparotomy, via direct laparoscopy, transabdominally, or percutaneously. The enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can be administered by percutaneous injection into the renal cortex of the kidney, or can be administered by percutaneous insertion of a catheter to puncture the renal capsule and then injecting the enriched heterogeneous renal cell population into the kidney. The enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can be administered by injection into the renal cortex of one or both kidneys of a patient. Administration of the enriched heterogeneous renal cell population or a pharmaceutical composition comprising the enriched heterogeneous renal cell population can be carried out by two injections, wherein the first injection is into the renal cortex of one kidney of the patient and the second injection is into the renal cortex of the other kidney of the patient.

[0049] An enriched heterogeneous renal cell population or a pharmaceutical composition comprising an enriched heterogeneous renal cell population is administered in a therapeutically effective dose by any suitable route. For a therapeutically effective dose or amount to be administered to a patient in need of treatment for a kidney disease, it may include about 1 - 9 x 10 6 cells of the enriched heterogeneous renal cell population per gram of the estimated patient kidney weight. A therapeutically effective amount of the pharmaceutical composition may be about 1.0×10 6 , about 2.0×10 6 , about 3.0×10 6 , about 4.0×10 6 , about 5.0×10 6 , about 6.0×10 6 , about 7.0×10 6 , about 8.0×10 6 , about 9.0×10 6 , about 2.0 - 7.0×10 6 , about 4.0 - 7.0×10 6 between or about 5.0×10 6 -7.0×10 6 between cells of the enriched heterogeneous renal cell population per gram of the estimated patient kidney weight.

[0050] In a method of treating a kidney disease, the administration of a therapeutic composition comprising a heterogeneous renal cell population to a patient can be by a first and a second injection. The first and second injections can be administered at intervals of about 3 to 12 months. The first and second injections can be administered at intervals of about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 12 months. The first and second injections can be administered at intervals between about 3 to 6 months, between about 6 to 9 months, between about 9 to 12 months, between about 3 to 9 months, between about 6 to 9 months, or between about 6 to 12 months.

[0051] Those skilled in the art will recognize or be able to determine many equivalents of the specific embodiments described herein using only routine experimentation. These equivalents are intended to be covered by the appended claims.

[0052] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the extent that each individual publication, patent, or patent application is specifically and individually indicated to be incorporated by reference in its entirety. Examples

[0053] Example 1 - Selected renal cells (SRC) stabilize and improve renal function in part by co - expressing a unique set of markers involved in kidney development

[0054] Introduction. Selected renal cells (SRC) are examples of enriched heterogeneous renal cell populations enriched for renal epithelial cells. SRC is being advanced as an autologous cell - based therapy for chronic kidney disease (CKD). A study combining empirical SRC gene expression data with gene ontology was conducted to determine whether human SRC, as an independent platform, restores diseased CKD renal function and potentially initiates new kidney development, at least in part, by co - expressing proteins involved in kidney development.

[0055] Methods / Human SRC Nodes: Proteomic analysis showed that a human enriched heterogeneous renal cell population, SRC, co - expresses eight markers, Six2, Osr1, Lhx1, Ret, Fgf8, Rack1, Nphs1, and Nphs2 (Stavas J, et al., KI Reports, in press). These eight markers were selected as the nodes to be analyzed in this study. Co - expression data for each of the eight markers are provided in Table 1.

[0056] Table 1: Compiled data for fluorescence - activated cell sorting (FACS) analysis

[0057]

[0058] Methods / Co - expression analysis. The eight nodes co - expressed by SRC, e.g., an example enriched heterogeneous renal cell population, were converted to their gene symbols (Homo sapiens) viz. six2, osr1, lhx1, ret, fgf8, rack1 / gnb2l1, nphs1, and nphs2 and input into STRING (string - db.org, last accessed 06 / 06 / 2022) and Genemania (https: / / genemania.org, last accessed 06 / 06 / 2022) to query their co - expression in human tissues.

[0059] Methods / Renal progenitor cell tracing: To map the expression of the eight markers in cells involved in kidney development, their gene symbols were queried using the Human fetal Kidney Atlas - Semraulab (leidenuniv.nl, last accessed 06 / 06 / 2022). Violin plots were generated to report the expression levels of the eight markers in up to 22 cell types.

[0060] Method / human SRC interactome and biological process analysis: An interactome of human SRC was constructed by inputting the gene symbols of eight nodes co-expressed with human SRC into Genemania and querying for genetic interactions. Biological process maps were created using ShinyGO (bioinformatics.sdstate.edu / go / http: / / bioinformatics.sdstate.edu / go / , last accessed 06 / 03 / 2022; false discovery rate cutoff = 0.05; number of pathways shown # = 40) and Humanbase (https: / / hb.flatironisntiute.org; last accessed 06 / 04 / 2022; biological process gene network; default for minimum interaction confidence = 0.1 and default for maximum number of genes = 15).

[0061] Method for scRNA-seq to map kirrel1 expression: The SRC was submitted to scRNA-seq to map gene expression. Briefly, the SRC aliquots were stained with acridine orange and propidium iodide, and the viability, concentration, and singleness of the cells were evaluated using a LUNA-FX7 dual fluorescence cell counter (Logos Biosystems). Then, the cells were processed using a 10x Genomics Chromium Controller and Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3.1 Dual Index Kit (PN-1000268) according to the manufacturer's user guide (https: / / tinyurl.com / 4855859x). Briefly, approximately 5,000 cells from each sample were loaded onto a chromium chip G, with a target recovery of 3,000 cells per sample for library preparation. Single cells, reverse transcription reagents, and gel beads coated with barcoded oligomers were encapsulated together in oil droplets to generate emulsion gel beads (GEMs). Reverse transcription was performed using a C1000 thermal cycler (Bio-Rad) to generate a complementary DNA (cDNA) library labeled with cell barcodes and unique molecular identifiers (UMIs). The GEMs were disrupted and the cDNA was purified using Dynabeads MyOne SILANE beads (Invitrogen), followed by 12 cDNA amplification cycles. The amplified cDNA library was purified using SPRIselect magnetic beads (Beckman Coulter) and quantified using an Agilent Bioanalyzer high-sensitivity DNA chip (Agilent Technologies). Then, fragmentation, end repair, A-tailing, and dual-sided size selection were performed using SPRIselect beads. Illumina-compatible adapters were ligated to the size-selected cDNA fragments. Then, the adapter-ligated cDNA was purified using SPRIselect beads. During 12 amplification cycles, uniquely identifiable indices were added to this cDNA. Then, the completed sequencing library was purified using SPRIselect beads, visualized using a Bioanalyzer high-sensitivity DNA chip, and mixed in equimolar ratios. The pooled libraries were sequenced on a NextSeq 2000 machine (Illumina) at the UNC High-Throughput Sequencing Facility. The pooled libraries were denatured and diluted according to the standard Illumina protocol, 1% PhiX sequencing control (Illumina) was incorporated, and sequencing was performed in paired-end format (read 1: 28 cycles, i7 index: 10 cycles, i5 index: 10 cycles, read 2: 90 cycles) on one P3 flow cell, with a total depth of 1.2 billion read pairs passing the quality filter.Library demultiplexing and initial analysis were performed using Cell Ranger 7.1.0 with default settings, and analysis was performed using 10X Genomics' Loupe Browser.

[0062] Results / Analysis of SRC: Query two knowledge bases, STRING and Genemania, to determine whether the co-expression of six2, osr1, lhx1, ret, fgf8, rack1, nphs1, and nphs2 has been reported in human tissues. Inputting these eight genes into either database returns a network with fewer rack1 and ret ( Figure 1A and 1B ), indicating that not all eight nodes are known to be co-expressed in human tissues.

[0063] To map whether the eight markers co-expressed by SRC (e.g., the example-enriched heterogeneous renal cell population) are expressed by cells involved in early kidney development, query the eight markers in the human fetal kidney atlas. The human fetal kidney atlas is a repository of single-cell transcriptomes related to 22 cell types involved in human kidney development. Querying this library indicates that the human SRC markers are differentially expressed by multiple renal progenitor cells, including epithelial and mesenchymal progenitor cell pools. The gene six2 ( Figure 2A ), osr1 ( Figure 2B ), and lhx1 ( Figure 2C ) are mainly expressed by nephron progenitor cells, and the gene lhx1 was also observed to be expressed in renal vesicles, comma-shaped bodies, S-shaped bodies, podocyte progenitor cells, connecting tubules, and Henle's loop cells. The expression of ret ( Figure 2D ) occurs in connecting tubule cells and ureteric buds, while the expression of fgf8 ( Figure 2E ) is observed in nephron progenitor cells, pre-tubular aggregate cells, and comma-shaped body cells. The expression of Rack1 / guanine nucleotide-binding protein subunit beta-2-like 1 (gnb2l) ( Figure 2F ) is relatively uniform among multiple cell types, including the above cells and podocytes, mesangial cells, and endothelial cells. Nphs1 ( Figure 2G ) and nphs2 ( Figure 2H ) are expressed on podocyte progenitor cells and podocytes. These findings suggest that SRC, as a single platform, may represent multiple renal progenitor cells involved in the spatio-temporal patterning of the kidney.

[0064] Results / Human SRC interactome: Gene ontology was used to construct the human SRC interactome by mapping the interactions of the eight markers (e.g., nodes) co-expressed by SRC. In addition to the eight nodes, the human SRC interactome ( Figure 3) may include other major nodes, including Kirre-like nephrosis protein family adhesion molecule 1 (kirrel 1). The kirrel 1 gene product NEPH1 is a member of the nephrin-like protein family and interacts with podocin to maintain podocyte barrier integrity; mutations in kirrel1 are known to be associated with nephrotic syndrome (Solanki AK, et al., Kidney Int, 96(2019):883-889).

[0065] Gene ontology biological process analysis showed that components within the human SRC interactome play an integral role in all aspects of early kidney development, including ureteric bud induction, mesenchymal cell differentiation, metanephric glomerular development, anterior renal tubule aggregation, and the formation of comma and S-shaped bodies, etc. ( Figure 4 ). The roles of components in the human SRC interactome obtained in the gene ontology biological process analysis are consistent with the results obtained from the above SRC analysis. That is, SRC, as a single platform, covers components that are typically expressed by renal progenitor cells.

[0066] Finally, Humanbase was used to investigate other kidney processes that may be related to the human SRC interactome. HumanBase applies machine learning algorithms to learn biological associations from large genomic data sets. Its comprehensive analysis goes beyond the existing biological knowledge reflected in the literature to identify new, data-driven associations. HumanBase analysis showed that angiogenesis is likely to be related to the SRC interactome, as evidenced by the robust network strength of this process ( Figure 5A ; network strength 0.45). Consistent with the findings of Humanbase, analysis of the human SRC secretome showed elevated levels of VEGF-A (Stavas J, et al., KI Reports, in press). Na+ transport emerged as another process that is likely to be related to the SRC network ( Figure 5B ; network strength 0.74). Sodium transport in distal and connecting tubule cells occurs via the Na + -Cl − cotransporter, which is expressed in the developing kidney and only in the cells of the neonatal distal tubule (Hadchouel J and Jeunemaitre X, Cell Metab 4(2006):335-7).

[0067] Results / scRNAseq: To further confirm that SRC includes cells expressing kirrel1, SRC was submitted to scRNAseq and probed for kirrel1 expression. scRNAseq data confirmed that SRC expresses kirrel1. See Tables 6 and 2.

[0068] Table 2: Kirrel1 expression

[0069]

[0070] SRC gene expression data elucidate the mechanism by which SRC may mediate the regenerative and restorative activities of diseased kidneys. The expression of six2, osr1, lhx1, ret, fgf8, rack1, nphs1, and nphs2 genes has identified SRC as a cell that may affect the signaling cascade mediating kidney development or kidney regeneration; SRC includes cells that express genes also expressed by kidney (e.g., nephron) progenitor cells. The identification of kirrel1 expression in SRC also suggests that the biological activity of SRC may be achieved by affecting or participating in glomerular barrier function. In summary, empirical gene expression data combined with interactome analysis support SRC (an enriched heterogeneous renal cell population) as an independent platform for the treatment of CKD; support SRC as a therapeutic modality that, consistent with its clinical activity, has the mechanistic ability to restore endogenous repair and functional maintenance of diseased CKD kidneys, as well as the potential to initiate new kidney development.

Claims

1. A method for identifying an enriched heterogeneous renal cell population with therapeutic potential, the method comprising: Determining whether the cells of the enriched heterogeneous renal cell population express: (i) NEPH1; and (ii) one or more markers, wherein the one or more markers comprise one or more of the following: nephrin, podocin, sine oculis homeobox homolog 2 (SIX2), odd skipped related 1 (OSR1), LIM homeobox protein 1 (LHX1), fibroblast growth factor 8 (FGF8), receptor for activated C kinase 1 (RACK1), or rearranged during transfection (RET); and If it is determined that the cells of the enriched heterogeneous renal cell population express: (i) NEPH1; and (ii) the one or more markers, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

2. The method according to claim 1, wherein the one or more markers comprise SIX2.

3. The method according to claim 1, wherein the one or more markers comprise OSR1.

4. The method according to claim 1, wherein the one or more markers comprise LHX1.

5. The method according to claim 1, wherein the one or more markers comprise FGF8.

6. The method according to claim 1, wherein the one or more markers comprise RACK1.

7. The method according to claim 1, wherein the one or more markers comprise RET.

8. The method according to claim 1, wherein the one or more markers comprise podocin.

9. The method according to claim 1, wherein the one or more markers comprise nephrin.

10. The method according to claim 1, wherein the one or more markers comprise at least two markers, wherein the at least two markers comprise: (a) SIX2 and OSR1; (b) SIX2 and LHX1; (c) SIX2 and RET; (d) SIX2 and FGF8; (e) OSR1 and LHX1; (f) OSR1 and RET; (g) OSR1 and FGF8; (h) LHX1 and RET; (i) LHX1 and FGF8; (j) RET and FGF8; (k) nephrin and SIX2; (l) nephrin and OSR1; (m) nephrin and LHX1; (n) nephrin and RET; (o) nephrin and FGF8; (p) podocin and nephrin; (q) podocin and SIX2; (r) podocin and OSR1; (s) podocin and LHX1; (t) podocin and RET; (u) podocin and FGF8; (v) RACK1 and nephrin; (w) RACK1 and podocin; (x) RACK1 and SIX2; (y) RACK1 and OSR1; (z) RACK1 and LHX1; (aa) RACK1 and RET; or (bb) RACK1 and FGF8.

11. The method according to claim 1, wherein the one or more markers comprise at least three markers, and the at least three markers comprise: (a) SIX2, OSR1, and LHX1; (b) SIX2, OSR1, and RET; (c) SIX2, OSR1, and FGF8; (d) SIX2, LHX1, and RET; (e) SIX2, LHX1, and FGF8; (f) SIX2, RET, and FGF8; (g) OSR1, LHX1, and RET; (h) OSR1, LHX1, and FGF8; (i) OSR1, RET, and FGF8; (j) LHX1, RET, and FGF8; (k) Nephrin, SIX2, and OSR; (l) Nephrin, SIX2, and LHX1; (m) Nephrin, SIX2, and RET; (n) Nephrin, SIX2, and FGF8; (o) Nephrin, OSR, and LHX1; (p) Nephrin, OSR, and RET; (q) Nephrin, OSR, and FGF8; (r) Nephrin, LHX1, and RET; (s) Nephrin, LHX1, and FGF8; (t) Nephrin, RET, and FGF8; (u) Podocin, SIX2, and OSR; (v) Podocin, SIX2, and LHX1; (w) Podocin, SIX2, and RET; (x) Podocin, SIX2, and FGF8; (y) Podocin, OSR, and LHX1; (z) Podocin, OSR, and RET; (aa) Podocin, OSR, and FGF8; (bb) Podocin, LHX1, and RET; (cc) Podocin, LHX1, and FGF8; (dd) Podocin, RET, and FGF8; (ee) RACK1, SIX2, and OSR; (ff) RACK1, SIX2, and LHX1; (gg) RACK1, SIX2, and RET; (hh) RACK1, SIX2, and FGF8; (ii) RACK1, OSR, and LHX1; (jj) RACK1, OSR, and RET; (kk) RACK1, OSR, and FGF8; (ll) RACK1, LHX1, and RET; (mm) RACK1, LHX1, and FGF8; (nn) RACK1, RET, and FGF8; (oo) Nephrin, Podocin, and RACK1; (pp) Nephrin, Podocin, and SIX2; (qq) Nephrin, Podocin, and OSR; (rr) Nephrin, Podocin, and RET; (ss) Nephrin, Podocin, and LHX1; (tt) Nephrin, Podocin, and FGF8; (uu) Nephrin, RACK1, and SIX2; (vv) Nephrin, RACK1, and OSR; (ww) Nephrin, RACK1, and RET; (xx) Nephrin, RACK1, and LHX1; (yy) Nephrin, RACK1, and FGF8; (zz) Podocin, RACK1, and SIX2; (aaa) Podocin, RACK1, and OSR; (bbb) Podocin, RACK1, and RET; or (ccc) Podocin, RACK1, and LHX1;or (ddd) podocin, RACK1, and FGF8.; 12. The method according to claim 1, wherein the one or more markers comprise at least four markers, and the at least four markers comprise: (a) SIX2, OSR1, LHX1, and RET; (b) SIX2, OSR1, LHX1, and FGF8; (c) SIX2, LHX1, RET, and FGF8; (d) SIX2, OSR1, RET, and FGF8; (e) OSR1, LHX1, RET, and FGF8; (f) Nephrin, SIX2, OSR1, and LHX1; (g) Nephrin, SIX2, OSR1, and RET; (h) Nephrin, SIX2, OSR1, and FGF8; (i) Nephrin, SIX2, LHX1, and RET; (j) Nephrin, SIX2, LHX1, and FGF8; (k) Nephrin, SIX2, RET, and FGF8; (l) Nephrin, OSR1, LHX1, and FGF8; (m) Nephrin, OSR1, LHX1, and RET; (n) Nephrin, OSR1, RET, and FGF8; (o) Nephrin, LHX1, RET, and FGF8; (p) Nephrin, SIX2, OSR1, and LHX1; (q) Podocin, SIX2, OSR1, and RET; (r) Podocin, SIX2, OSR1, and FGF8; (s) Podocin, SIX2, LHX1, and RET; (t) Podocin, SIX2, LHX1, and FGF8; (u) Podocin, SIX2, RET, and FGF8; (v) Podocin, OSR1, LHX1, and FGF8; (w) Podocin, OSR1, LHX1, and RET; (x) Podocin, OSR1, RET, and FGF8; (y) Podocin, LHX1, RET, and FGF8; (z) RACK1, SIX2, OSR1, and FGF8; (aa) RACK1, SIX2, LHX1, and RET; (bb) RACK1, SIX2, LHX1, and FGF8; (cc) RACK1, SIX2, RET, and FGF8; (dd) RACK1, OSR1, LHX1, and FGF8; (ee) RACK1, OSR1, LHX1, and RET; (ff) RACK1, OSR1, RET, and FGF8; (gg) RACK1, LHX1, RET, and FGF8; (hh) Nephrin, Podocin, SIX2, and OSR1; (ii) Nephrin, Podocin, SIX2, and LHX1; (jj) Nephrin, Podocin, SIX2, and RET; (kk) Nephrin, Podocin, SIX2, and FGF8; (ll) Nephrin, Podocin, OSR1, and LHX1; (mm) Nephrin, Podocin, OSR1, and RET; (nn) Nephrin, Podocin, OSR1, and FGF8; (oo) Nephrin, Podocin, LHX1, and RET; (pp) Nephrin, Podocin, LHX1, and FGF8; (qq) Nephrin, Podocin, RET, and FGF8; (rr) RACK1, Podocin, SIX2, and OSR1;(ss)RACK1, podocin, SIX2, and LHX1; (tt)RACK1, podocin, SIX2, and RET; (uu)RACK1, podocin, SIX2, and FGF8; (vv)RACK1, podocin, OSR1, and LHX1; (ww)RACK1, podocin, OSR1, and RET; (xx)RACK1, podocin, OSR1, and FGF8; (yy)RACK1, podocin, LHX1, and RET; (zz)RACK1, podocin, LHX1, and FGF8; (aaa)RACK1, podocin, RET, and FGF8; (bbb)RACK1, nephrocystin, SIX2, and OSR1; (ccc)RACK1, nephrocystin, SIX2, and LHX1; (ddd)RACK1, nephrocystin, SIX2, and RET; (eee)RACK1, nephrocystin, SIX2, and FGF8; (fff)RACK1, nephrocystin, OSR1, and LHX1; (ggg)RACK1, nephrocystin, OSR1, and RET; (hhh)RACK1, nephrocystin, OSR1, and FGF8; (iii)RACK1, nephrocystin, LHX1, and RET; (jjj)RACK1, nephrocystin, LHX1, and FGF8; (kkk)RACK1, nephrocystin, RET, and FGF8; (lll)RACK1, nephrocystin, podocin, and SIX2; (mmm)RACK1, nephrocystin, podocin, and RET; (nnn)RACK1, nephrocystin, podocin, and LHX1; (ooo)RACK, nephrocystin, podocin, and OSR1; or (ppp)RACK, nephrocystin, podocin, and FGF8.; 13. The method according to claim 1, wherein the one or more markers comprise at least five markers, and the at least five markers comprise: (a) SIX2, OSR1, LHX1, RET, and FGF8; (b) Nephrin, SIX2, OSR1, LHX1, and RET; (c) Nephrin, SIX2, OSR1, LHX1, and FGF8; (d) Nephrin, SIX2, LHX1, RET, and FGF8; (e) Nephrin, SIX2, OSR1, RET, and FGF8; (f) Nephrin, OSR1, LHX1, RET, and FGF8; (g) Podocin, SIX2, OSR1, LHX1, and RET; (h) Podocin, SIX2, OSR1, LHX1, and FGF8; (i) Podocin, SIX2, LHX1, RET, and FGF8; (j) Podocin, SIX2, OSR1, RET, and FGF8; (k) Podocin, OSR1, LHX1, RET, and FGF8; (l) RACK1, SIX2, OSR1, LHX1, and RET; (m) RACK1, SIX2, OSR1, LHX1, and FGF8; (n) RACK1, SIX2, LHX1, RET, and FGF8; (o) RACK1, SIX2, OSR1, RET, and FGF8; (p) RACK1, OSR1, LHX1, RET, and FGF8; (q) Nephrin, Podocin, SIX2, OSR1, and LHX1; (r) Nephrin, Podocin, SIX2, OSR1, and RET; (s) Nephrin, Podocin, SIX2, OSR1, and FGF8; (t) Nephrin, Podocin, SIX2, LHX1, and RET; (u) Nephrin, Podocin, SIX2, LHX1, and FGF8; (v) Nephrin, Podocin, SIX2, RET, and FGF8; (w) Nephrin, Podocin, OSR1, LHX1, and RET; (x) Nephrin, Podocin, OSR1, LHX1, and FGF8; (y) Nephrin, Podocin, OSR1, RET, and FGF8; (z) Nephrin, Podocin, LHX1, RET, and FGF8; (aa) Nephrin, RACK1, SIX2, OSR1, and LHX1; (bb) Nephrin, RACK1, SIX2, OSR1, and RET; (cc) Nephrin, RACK1, SIX2, OSR1, and FGF8; (dd) Nephrin, RACK1, SIX2, LHX1, and RET; (ee) Nephrin, RACK1, SIX2, LHX1, and FGF8; (ff) Nephrin, RACK1, SIX2, RET, and FGF8; (gg) Nephrin, RACK1, OSR1, LHX1, and RET; (hh) Nephrin, RACK1, OSR1, LHX1, and FGF8; (ii) Nephrin, RACK1, OSR1, RET, and FGF8; (jj) Nephrin, RACK1, LHX1, RET, and FGF8;(kk) Nephrin, RACK1, SIX2, OSR1, and LHX1; (ll) Nephrin, RACK1, SIX2, OSR1, and RET; (mm) Nephrin, RACK1, SIX2, OSR1, and FGF8; (nn) Nephrin, RACK1, SIX2, LHX1, and RET; (oo) Nephrin, RACK1, SIX2, LHX1, and FGF8; (pp) Nephrin, RACK1, SIX2, RET, and FGF8; (qq) Nephrin, RACK1, OSR1, LHX1, and RET; (rr) Nephrin, RACK1, OSR1, LHX1, and FGF8; (ss) Nephrin, RACK1, OSR1, RET, and FGF8; (tt) Nephrin, RACK1, LHX1, RET, and FGF8; (uu) Nephrocystin, Nephrin, RACK1, SIX2, and OSR1; (vv) Nephrocystin, Nephrin, RACK1, SIX2, and LHX1; (ww) Nephrocystin, Nephrin, RACK1, SIX2, and RET; (xx) Nephrocystin, Nephrin, RACK1, SIX2, and FGF8; (yy) Nephrocystin, Nephrin, RACK1, OSR1, and LHX1; (zz) Nephrocystin, Nephrin, RACK1, OSR1, and RET; (aaa) Nephrocystin, Nephrin, RACK1, OSR1, and FGF8; (bbb) Nephrocystin, Nephrin, RACK1, LHX1, and RET; (ccc) Nephrocystin, Nephrin, RACK1, LHX1, and FGF8; or (ddd) Nephrocystin, Nephrin, RACK1, RET, and FGF8.; 14. The method according to claim 1, wherein the one or more markers comprise at least six markers, and the at least six markers comprise: (a) Nephrin, SIX2, OSR1, LHX1, RET, and FGF8; (b) Podocin, SIX2, OSR1, LHX1, RET, and FGF8; (c) RACK1, SIX2, OSR1, LHX1, RET, and FGF8; (d) Nephrin, Podocin, SIX2, OSR1, LHX1, and RET; (e) Nephrin, Podocin, SIX2, OSR1, LHX1, and FGF8; (f) Nephrin, Podocin, SIX2, LHX1, RET, and FGF8; (g) Nephrin, Podocin, SIX2, OSR1, RET, and FGF8; (h) Nephrin, Podocin, OSR1, LHX1, RET, and FGF8; (i) Nephrin, RACK1, SIX2, OSR1, LHX1, and RET; (j) Nephrin, RACK1, SIX2, OSR1, LHX1, and FGF8; (k) Nephrin, RACK1, SIX2, LHX1, RET, and FGF8; (l) Nephrin, RACK1, SIX2, OSR1, RET, and FGF8; (m) Nephrin, RACK1, OSR1, LHX1, RET, and FGF8; (n) Podocin, RACK1, SIX2, OSR1, LHX1, and RET; (o) Podocin, RACK1, SIX2, OSR1, LHX1, and FGF8; (p) Podocin, RACK1, SIX2, LHX1, RET, and FGF8; (q) Podocin, RACK1, SIX2, OSR1, RET, and FGF8; (r) Podocin, RACK1, OSR1, LHX1, RET, and FGF8; (s) Nephrin, Podocin, RACK1, SIX2, OSR1, and LHX1; (t) Nephrin, Podocin, RACK1, SIX2, OSR1, and RET; (u) Nephrin, Podocin, RACK1, SIX2, OSR1, and FGF8; (v) Nephrin, Podocin, RACK1, SIX2, LHX1, and RET; (w) Nephrin, Podocin, RACK1, SIX2, LHX1, and FGF8; (x) Nephrin, Podocin, RACK1, SIX2, RET, and FGF8; (y) Nephrin, Podocin, RACK1, OSR1, LHX1, and RET; (z) Nephrin, Podocin, RACK1, OSR1, LHX1, and FGF8; (aa) Nephrin, Podocin, RACK1, OSR1, RET, and FGF8; or (bb) Nephrin, Podocin, RACK1, LHX1, RET, and FGF8.

15. The method of claim 1, wherein the one or more markers comprise at least seven markers, wherein the at least seven markers comprise: (a) Nephrin, RACK1, SIX2, OSR1, LHX1, RET, and FGF8; (b) Nephrosis protein, RACK1, SIX2, OSR1, LHX1, RET, and FGF8; (c) Nephrosis protein, Nephrin, SIX2, OSR1, LHX1, RET, and FGF8; (d) Nephrosis protein, Nephrin, RACK1, OSR1, LHX1, RET, and FGF8; (e) Nephrosis protein, Nephrin, RACK1, SIX2, LHX1, RET, and FGF8; (f) Nephrosis protein, Nephrin, RACK1, SIX2, OSR1, RET, and FGF8; (g) Nephrosis protein, Nephrin, RACK1, SIX2, OSR1, LHX1, and FGF8; or (h) Nephrosis protein, Nephrin, RACK1, SIX2, OSR1, LHX1, and RET.

16. The method according to claim 15, wherein the at least seven markers comprise: nephrosis protein, nephrin, SIX2, OSR1, LHX1, RET, and FGF8.

17. The method according to claim 15, wherein the at least seven markers comprise: nephrosis protein, nephrin, RACK1, SIX2, OSR1, LHX1, and FGF8.

18. The method according to claim 15, wherein the at least seven markers comprise: nephrosis protein, nephrin, RACK1, SIX2, OSR1, LHX1, and FGF8.

19. The method according to any one of the preceding claims, wherein if it is determined that the cells of the enriched heterogeneous renal cell population express SIX2, OSR1, LHX1, RET, FGF8, RACK1, nephrosis protein, and nephrin, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

20. The method according to any one of claims 1-19, wherein the determination comprises determining the percentage of cells expressing NEPH1 in the enriched heterogeneous renal population.

21. The method according to claim 20, wherein if it is determined that about 2.0% to about 8.0% of the cells in the population express NEPH1, then the enriched heterogeneous renal cell population is identified as having therapeutic potential.

22. The method according to any one of claims 1-9, wherein the determination comprises determining the percentage of cells expressing the one or more markers in the enriched heterogeneous population.

23. The method according to claim 10, wherein the determination comprises determining the percentage of cells expressing the at least two markers in the enriched heterogeneous population.

24. The method according to claim 11, wherein the determination comprises determining the percentage of cells expressing the at least three markers in the enriched heterogeneous population.

25. The method according to claim 12, wherein the determination comprises determining the percentage of cells expressing the at least four markers in the enriched heterogeneous population.

26. The method according to claim 13, wherein the determination comprises determining the percentage of cells expressing the at least five markers in the enriched heterogeneous population.

27. The method according to claim 14, wherein said determining comprises determining the percentage of cells expressing said at least six markers in said enriched heterogeneous population.

28. The method according to any one of claims 15 - 18, wherein said determining comprises determining the percentage of cells expressing said at least seven markers in said enriched heterogeneous renal cell population.

29. The method according to any one of claims 22 - 28, wherein said determining further comprises determining the percentage of cells expressing NEPH1 in said enriched heterogeneous renal population.

30. The method according to claim 29, wherein if it is determined that about 2.0% to about 8.0% of the cells in said population express NEPH1, said enriched heterogeneous renal cell population is identified as having therapeutic potential.

31. The method according to any one of claims 1 - 30, further comprising determining whether the cells of said enriched heterogeneous renal cell population express lefty, and if it is further determined that the cells of said enriched heterogeneous renal cell population express lefty, identifying said enriched heterogeneous renal cell population as having therapeutic potential.

32. The method according to any one of claims 1 - 31, further comprising determining whether the cells of said enriched heterogeneous renal cell population express nodal, and if it is further determined that the cells of said enriched heterogeneous renal cell population express nodal, identifying said enriched heterogeneous renal cell population as having therapeutic potential.

33. The method according to any one of claims 1 - 32, further comprising determining whether the cells of said enriched heterogeneous renal cell population express CD24, and if it is further determined that the cells of said enriched heterogeneous renal cell population express CD24, identifying said enriched heterogeneous renal cell population as having therapeutic potential.

34. The method according to any one of the preceding claims, wherein said enriched heterogeneous renal cell population is prepared by a method comprising a density gradient separation step.

35. The method according to claim 34, wherein the enriched heterogeneous renal cell population prepared by said method comprises cells having a buoyant density greater than about 1.04 g / mL.

36. The method according to any one of claims 34 or 35, wherein said enriched heterogeneous renal cell population is hypoxia - tolerant and has been cultured under hypoxic conditions prior to said density gradient separation step.

37. A composition comprising an enriched heterogeneous renal cell population identified by the method according to any one of the preceding claims.

38. A method for treating kidney diseases, the method comprising: Administering a therapeutically effective amount of the composition according to claim 37.

Images