Methods for identifying neuripotent cells
View Patent ↗The invention is a method for using an avian embryo to identify the neuripotency of a cell population. The invention may be used for applications such as screening for candidate neuripotent cell lines for masterbanking, validation of working cell banks, and identifying agents and conditions capable of inducing neural differentiation in a cell population.
1. A method for identifying a neuripotent cell population comprising:
(a) obtaining a cell sample from a population of donor cells;
(b) contacting an intact ventricle in the neural tissue of an avian embryo with said cell sample; and
(c) identifying that at least a portion of cells in said ceil sample demonstrate a neuripotent phenotype after contacting the intact ventricle in the neural tissue of said avian embryo.
2. The method of claim 1 , wherein said neuripotent phenotype is selected from at least one of migration, integration, survival and penetration.
3. The method of claim 1 , wherein said population of donor cells comprises mammalian cells.
4. The method of claim 3 , wherein said mammalian cells are human cells.
5. The method of claim 4 , wherein said human cells are neural cells.
6. The method of claim 5 , wherein said neural cells are obtained from fetal tissue.
7. The method of claim 3 , wherein said mammalian cells are neural cells.
8. The method of claim 1 , wherein said avian embryo comprises a chicken embryo.
9. The method of claim 1 , wherein said intact ventricle is a lateral ventricle.
10. The method of claim 9 , wherein said lateral ventricle is in the telencephalon of said avian embryo.
11. The method of claim 1 , wherein said population of donor cells is free of immortalized cells.
12. The method of claim 1 , further comprising banking the population of donor cells.
13. The method of claim 12 , wherein said banking comprises masterbanking.
14. The method of claim 1 , wherein said method is used to confirm the neuripotency of a masterbank of neuripotent cells.
15. The method of claim 1 , wherein said method is used to confirm the neuripotency of a working bank of neuripotent cells.
16. The method of claim 1 , wherein said ventricle is in the brain of said avian embryo.
17. A method for measuring the activity of a neuripotent cell population comprising:
(a) obtaining a cell sample from said neuripotent cell population;
(b) contacting an intact ventricle in the neural tissue of an avian embryo with said cell sample;
(c) measuring the activity of said cell sample based on the ability of said cell sample to migrate, survive, integrate and/or penetrate said avian embryo.
18. The method of claim 17 , wherein said cell sample is treated with an activating agent.
19. The method of claim 17 , wherein said intact ventricle is a lateral ventricle.
20. The method of claim 17 , wherein said intact ventricle comprises germinal epithelium.
21. The method of claim 1 , wherein said intact ventricle comprises germinal epithelium.
22. The method of claim 17 , wherein said intact ventricle is a lateral ventricle in the telencephalon of said avian embryo.
23. A method for identifying a population of cells having a neural stem cell phenotype, said method comprising:
(a) providing a population of cells;
(b) providing an avian embryo comprising neural tissue having one or more intact ventricles;
(c) contacting said one or more intact ventricles with said population of cells; and
(d) observing at least one of migration, integration, survival and penetration by at least a portion of the contacted cells in said one or more intact ventricles;
(e) wherein said observing identities said population of cells as having a neural stem cell phenotype.
24. The method of claim 23 , wherein said population of cells comprises mammalian cells.
25. The method of claim 24 , wherein said mammalian cells are human cells.
26. The method of claim 25 , wherein said human cells are neural cells.
27. The method of claim 24 , wherein said mammalian cells are neural cells.
28. The method of claim 23 , wherein said avian embryo is a chicken embryo.
29. The method of claim 23 , wherein contacting comprises contacting said population of cells with an intact lateral ventricle.
30. The method of claim 23 , wherein said population of cells is obtained from a donor source selected from at least one of tissue, cultured cells, a clonal cell line, and a primary culture of cells.
31. The method of claim 30 , further comprising creating a masterbank of cells from said donor source.
32. The method of claim 31 , wherein said donor source comprises neural tissue.
33. The method of claim 23 , wherein said population of cells is obtained from a masterbank of neural stem cells.
34. The method of claim 23 , wherein said population of cells is obtained from a working bank of neural stem cells.
35. The method of claim 23 , wherein contacting comprises contacting said population of cells with the germinal epithelium of said one or more intact ventricles.
36. The method of claim 23 , wherein said one or more intact ventricles are in the brain of said avian embryo.
37. A method for identifying a donor source for producing a masterbank of neural stem cells, said method comprising:
(a) providing a cell sample from a donor source;
(b) providing an avian embryo;
(c) contacting at least one intact ventricle in the brain of said avian embryo with said cell sample;
(d) observing in said at least one intact ventricle at least one of migration, integration, survival and penetration by at least a portion of the cells in said cell sample, wherein said observing identifies said donor source as a source of neural stem cells; and
(e) producing a masterbank of cells from said donor source.
38. The method of claim 37 , wherein said at least one ventricle is in at least one of the metencephalon, mesencephalon, myelencephalon, diencephalon or telenchephalon of said brain of said avian embryo.
39. The method of claim 38 , wherein said at least one ventricle is a lateral ventricle in the telencephalon of said avian embryo.