Placental stem cells
View Patent ↗The present invention provides a method of extracting and recovering embryonic-like stem cells, including, but not limited to pluripotent or multipotent stem cells, from an exsanguinated human placenta. A placenta is treated to remove residual umbilical cord blood by perfusing an exsanguinated placenta, preferably with an anticoagulant solution, to flush out residual cells. The residual cells and perfusion liquid from the exsanguinated placenta are collected, and the embryonic-like stem cells are separated from the residual cells and perfusion liquid. The invention also provides a method of utilizing the isolated and perfused placenta as a bioreactor in which to propagate endogenous cells, including, but not limited to, embryonic-like stem cells. The invention also provides methods for propagation of exogenous cells in a placental bioreactor and collecting the propagated exogenous cells and bioactive molecules therefrom.
1. Isolated human CD34 − , CD10 + , SH2 + , CD90 + placental multipotent cells, wherein said cells are obtained from a placenta that has been drained of cord blood.
2. The isolated cells of claim 1 , wherein said cells are CD34 − , CD38 − , CD45 − , CD10 + , CD29 + , CD44 + , CD54 + , CD90 + , SH2 + , SH3 + , SH4 + and OCT-4 + .
3. The isolated cells of claim 1 that expand in culture medium comprising Dulbecco's Modified Eagle's Medium (DMEM) or Mesenchymal Stem Cell Growth Medium (MSCGM).
4. The isolated cells of claim 3 , wherein said culture medium comprises beta-mercaptoethanol.
5. The isolated cells of claim 3 , wherein said culture medium comprises an antibiotic.
6. The isolated cells of claim 1 , wherein said cells expand in serum-free medium.
7. The isolated cells of claim 1 that expand on an uncoated surface.
8. An isolated homogeneous cell population comprising the isolated cells of claim 1 .
9. An isolated homogeneous cell population comprising the isolated cells of claim 2 .
10. A cell culture comprising the cell population of claim 8 .
11. The isolated cells of claim 1 , wherein said cells have been genetically engineered to produce a protein of interest.
12. The isolated cells of claim 1 , wherein said cells are primary isolates.
13. The isolated cells of claim 1 that have undergone at least one doubling in culture.
14. An isolated population of human CD34 − placental multipotent cells wherein said population is one or more of CD29 + , CD45 − , CD90 + , SH2 + , SH3 + , SH4 + , or MHC Class II − , wherein said cells adhere to tissue culture plastic, and wherein said cells have been obtained from a placenta that has been drained of cord blood and perfused to remove residual blood.
15. The isolated population of claim 14 , wherein said population is CD34 − , SH2 + , SH3 + and SH4 + .
16. The isolated population of claim 14 , wherein said population is CD34 − and MHC Class II − .
17. The isolated population of claim 14 , wherein said population is CD29 + , CD34 − , CD45 − , CD90 + , SH2 + , SH3 + , SH4 + , and MHC Class II − .
18. The isolated population of claim 14 wherein said population has the potential to differentiate into cells of an osteogenic phenotype.
19. The isolated population of claim 14 comprising cells that are fetal in origin.
20. The isolated population of claim 14 that expands in culture medium comprising DMEM or Mesenchymal Stem Cell Growth Medium (MSCGM).
21. The isolated population of claim 14 that expands in culture medium comprising fetal bovine serum (FBS).
22. The isolated population of claim 21 , wherein said culture medium comprises beta-mercaptoethanol.
23. The isolated population of claim 21 , wherein said culture medium comprises an antibiotic.
24. The isolated population of claim 14 that expands on an uncoated surface.
25. An isolated homogeneous cell population comprising the population of claim 14 .
26. An isolated homogeneous cell population comprising the population of claim 15 .
27. An isolated homogeneous cell population comprising the population of claim 16 .
28. An isolated homogeneous cell population comprising the population of claim 17 .
29. A cell culture comprising the population of claim 14 .
30. The isolated cells of claim 8 that have undergone at least one doubling in culture.
31. The isolated cells of claim 9 that have undergone at least one doubling in culture.