CELL POTENCY ASSAY
Provided herein are methods for assessing cell potency.
1 . A method for assessing cell potency, said method comprising the following steps (i) obtaining a population of cells; (ii) culturing the cells for a time period that minimizes variability in cell number once the cells have been cultured for the time period; and (iii) performing an assay to determine whether the cells produce a marker that acts as a surrogate for potency of the cells.
2 . The method of claim 1 , wherein the time period in step (ii) is about 48 hours.
3 . The method of claim 2 , wherein the marker that acts as a surrogate for potency of the cells is prostaglandin E2 (PGE2), wherein expression of PGE2 is a surrogate for potency of the cells.
4 . A method for assessing cell potency, said method comprising the following steps (i) obtaining a population of cells; (ii) culturing the cells for a time period that minimizes variability in cell number once the cells have been cultured for the time period; (iii) further culturing the cells in a medium that comprises one or more agents capable of inducing production of one or more genes in the cells; and (iv) performing an assay to determine whether the cells produce a marker that acts as a surrogate for potency of the cells.
5 . The method of claim 4 , wherein the time period in step (ii) is about 48 hours.
6 . The method of claim 5 , wherein the marker that acts as a surrogate for potency of the cells is PGE2, wherein expression of PGE2 is a surrogate for potency of the cells.
7 . The method of claim 6 , wherein said agent capable of inducing production of PGE2 by the cells is interleukin-1 (IL-1) beta.
8 . The method of claim 1 , wherein the population of cells is suitable for use as a cell therapy.
9 . The method of claim 1 , wherein the population of cells comprises mesenchymal stem cells, bone marrow-derived mesenchymal stem cells (BM-MSCs), tissue culture plastic-adherent CD34 − , CD10 + , CD105 + , CD200 + placental stem cells, embryonic stem cells, embryonic germ cells, induced pluripotent stem cells, mesenchymal stem cells, bone marrow-derived mesenchymal stem cells, bone marrow-derived mesenchymal stromal cells, tissue plastic-adherent placental stem cells (PDACs), umbilical cord stem cells, amniotic fluid stem cells, amnion derived adherent cells (AMDACs), osteogenic placental adherent cells (OPACs), adipose stem cells, limbal stem cells, dental pulp stem cells, myoblasts, endothelial progenitor cells, neuronal stem cells, exfoliated teeth derived stem cells, hair follicle stem cells, dermal stem cells, parthenogenically derived stem cells, reprogrammed stem cells, amnion derived adherent cells, or side population stem cells.
10 . The method of claim 1 , wherein the cells are human cells.
11 . The method of claim 1 , wherein the population of cells comprises placental stem cells.
12 . The method of claim 11 , wherein said placental cells adhere to tissue culture plastic and are CD34 − , CD10 + , CD105 + and CD200 + , as detectable by flow cytometry.
13 . The method of claim 1 , wherein the population of cells is obtained from a lot of cells that had been previously cryopreserved.
14 . The method of claim 1 , wherein the time period that minimizes variability in cell number is greater than 24 hours.
15 .- 17 . (canceled)
18 . The method of claim 1 , wherein the marker that acts as a surrogate for potency of the cells in the population of cells correlates with immunosuppressive activity of the cells.
19 . (canceled)
20 . The method of claim 1 , wherein the marker that acts as a surrogate for potency of the cells in the population of cells is ANG, EGF, ENA-78, FGF2, Follistatin, G-CSF, GRO, HGF, IL-6, IL-8, Leptin, MCP-1, MCP-3, PDGFB, PLGF, Rantes, TGFB1, Thrombopoietin, TIMP1, TIMP2, uPAR, VEGF, VEGFD, angiopoietin-1, angiopoietin-2, PECAM-1 (CD31; platelet endothelial cell adhesion molecule), laminin and/or fibronectin.
21 . The method of claim 1 , wherein said marker that acts as a surrogate for cell potency is detected by ELISA, by a MulitplexBead Assay, or by an assay that measures gene expression.
22 .- 24 . (canceled)
25 . The method of claim 1 , further comprising collection of the conditioned media from the cell culture.
26 . The method of claim 1 , wherein if the cells produce a marker that acts as a surrogate for potency of the cells, the original non-cultured population of cells is divided into a lot of cells suitable for administration to human subjects.
27 . (canceled)
28 . The method of claim 1 , wherein the population of cells is from a cryopreserved lot of cells.
29 .- 31 . (canceled)