Renal cell populations and uses thereof
The present invention concerns enriched heterogeneous mammalian renal cell populations characterized by biomarkers, and methods of making and using the same.
1. A method of identifying an enriched heterogeneous renal cell population as suitable for eliciting a regenerative response said method comprising:
contacting a sample of cells of an enriched heterogeneous renal cell population with labeled detection moieties, wherein each labeled detection moiety is directed to a different biomarker and is labeled with a different label, wherein biomarkers detected are gamma-glutamyl transpeptidase (GGT)-1, a cytokeratin (CK), vascular endothelial growth factor (VEGF), and kidney injury molecule-1 (KIM-1);
determining percentage of cells in the sample that express GGT-1 and the CK;
determining whether the sample comprises cells that secrete VEGF and KIM-1 in culture medium; and
identifying the enriched heterogeneous renal cell population as suitable for eliciting a regenerative response if it is determined that: (i) greater than 10% of cells of the sample express GGT-1, (ii) greater than 80% of cells of the sample express the CK and (iii) the sample comprises cells that secrete VEGF and KIM-1 in the culture medium.
2. The method of claim 1 wherein the biomarkers detected further comprise one or more of: AQP1, AQP2, AQP4, Calbindin, Calponin, CD117, CD133, CD146, CD24, CD31 (PECAM-1), CD54 (ICAM-1), CD73, CK18, CK19, CK40 to 67, CK7, CK8, CK8, CK18, CK19, combinations of CK8, CK18, CK19, Connexin 43, Cubilin, CXCR4 (Fusin), DBA, E-cadherin (CD324), EPO (erythropoeitin), GLEPP1 (glomerular epithelial protein 1), Haptoglobin, Itgb1 (Integrin (β1), MAP-2 (microtubule-associated protein 2), Megalin, N-cadherin, Nephrin, NKCC (Na—K—Cl-cotransporters), OAT-1 (organic anion transporter 1), Osteopontin, Pan-cadherin, PCLP1 (podocalyxin-like 1 molecule), Podocin, SMA (smooth muscle alpha-actin), Synaptopodin, THP (tamm-horsfall protein), Vimentin, αGST-1 (alpha glutathione S-transferase).
3. The method of claim 1 wherein the labeled detection moieties for biomarkers GGT-1 and CK are antibodies.
4. The method of claim 1 , wherein the enriched heterogeneous renal cell population is identified as suitable for eliciting a regenerative response if greater than 18% of cells of the sample express GGT-1.
5. The method of claim 4 , wherein the enriched heterogeneous renal cell population is identified as suitable for eliciting a regenerative response if greater than 85% of cells of the sample express the CK.
6. The method of claim 5 , wherein the CK is CK18.
7. The method of claim 6 , wherein the biomarkers detected further comprise AQP2.
8. The method of claim 7 , wherein the enriched heterogeneous renal cell population is identified as suitable for eliciting a regenerative response if between 3.0% and 53.7% of cells of the sample express AQP2.
9. The method of claim 8 , further comprising determining whether cells of the sample metabolize the PrestoBlue™ cell reagent; and identifying the enriched heterogeneous renal cell population as suitable for eliciting a regenerative response if it is determined that cells of the sample metabolize the PrestoBlue™ cell viability reagent.
10. The method of claim 1 , wherein the enriched heterogeneous renal cell population is identified as suitable for eliciting a regenerative response if greater than 85% of cells of the sample express the CK.
11. The method of claim 10 , wherein the CK is CK18.
12. The method of claim 1 , wherein the biomarkers detected further comprise AQP2.
13. The method of claim 12 , wherein the enriched heterogeneous renal cell population is identified as suitable for eliciting a regenerative response if between 3.0% and 53.7% of cells of the sample express AQP2.
14. The method of claim 1 , further comprising determining whether cells of the sample metabolize the PrestoBlue™ cell viability reagent; and identifying the enriched heterogeneous renal cell population as suitable for eliciting a regenerative response if it is determined that cells of the sample metabolize the PrestoBlue™ cell viability reagent.
15. The method of claim 1 , wherein the enriched heterogeneous renal cell population is derived from a mammalian kidney sample and is enriched for renal epithelial cells.
16. The method of claim 15 , wherein the mammalian kidney sample is a human kidney sample.