Markers capable of distinguishing cell pluripotency and uses thereof
Provided herein are compositions, methods and uses that relate to or result from the identification of markers that can distinguish between cells at different stages of pluripotency. Certain embodiments provide markers that can distinguish between parental cells (i.e. differentiated cells), partially pluripotent (i.e. partially reprogrammed) and pluripotent (i.e. fully reprogrammed cells). Also provided here are uses of such differential markers, for example, in identification of cell potential, in diagnostics, in differential separation, and in creating efficient workflows that involve fewer steps and lesser time in identifying or separating a desired reprogrammed clone or cell line from a mixture of cells at various stages of pluripotency. In certain embodiments, the activity of these markers can be manipulated to influence cell potential for research or medical purposes.
1. A method for identifying differentiated cells or partially reprogrammed cells within a mixture of differentiated, partially reprogrammed and pluripotent cells, the method comprising:
contacting a mixture of differentiated, partially reprogrammed and pluripotent cells with a detectably labeled antibody wherein the detectably labeled antibody binds to differentiated cells or partially reprogrammed cells but does not bind to pluripotent cells;
detecting the distinguishably detectable labeled antibody bound to the differentiated or partially reprogrammed cells;
identifying the differentiated cells or the partially reprogrammed cells based on the detectably labeled antibody binding thereto and identifying the pluripotent cells based on a lack of the detectably labeled antibody binding thereto;
wherein the detectably labeled antibody binds to one of CD44, beta 2 microglobulin, endosialin, cadherin 11 or colectin 12 expressed on cells and bound cells in the mixture are identified as the differentiated cells or partially programmed cells.
2. The method according to claim 1 , wherein the cell is a mammalian cell.
3. The method according to claim 1 , wherein the detectably labeled antibody binds to CD44.
4. The method according to claim 1 , wherein the detectable label comprises a fluorescent dye, biotin, or live stain.
5. The method according to claim 1 , wherein the proportion of differentiated cells or partially reprogrammed cells to pluripotent cells in the mixture of cells is determined.
6. The method according to claim 1 , wherein the method is performed on a reprogrammed mixture of cells.
7. A method for determining the differentiation state of a cell, the method comprising:
contacting a cell with a detectably labeled antibody which binds a marker selected from the group consisting of CD44, beta 2 microglobulin, endosialin, cadherin 11 and colectin 12;
detecting the expression level of the marker in a cell,
wherein the expression of the marker identifies the cell as a differentiated or partially differentiated cell and the lack of expression of the marker identifies the cell as a pluripotent cell.
8. The method according to claim 7 , wherein the detectable label comprises a fluorescent dye, biotin, or live stain.
9. The method according to claim 7 , wherein the detectably labeled antibody binds to CD44.
10. The method according to claim 7 , wherein the cell is a mammalian cell.
11. The method according to claim 7 , wherein the cell had been subjected to a reprogramming workflow.