Placental stem cell populations

1 . A method of producing a cell population comprising identifying placental cells that adhere to a substrate and:

express CD200 and HLA-G;

express CD73, CD105, and CD200;

express CD200 and OCT-4;

express CD73, CD105 and HLA-G;

express CD73 and CD105, and facilitate the formation of one or more embryoid-like bodies in a population of placental cells when said population is cultured under conditions that allow for the formation of embryoid-like bodies; or

express OCT-4, and (c) facilitate the formation of one or more embryoid-like bodies in a population of placental cells when said population is cultured under conditions that allow for the formation of embryoid-like bodies; or any combination of the foregoing;

and isolating said cells from other cells to form a cell population.

2 . The method of claim 1 , wherein said substrate comprises fibronectin.

3 . The method of claim 1 , wherein said identifying is accomplished using an antibody.

4 . The method of claim 1 , wherein said identifying is accomplished using flow cytometry.

5 . The method of claim 1 , wherein said isolating is accomplished using magnetic beads.

6 . The method of claim 1 , wherein said isolating is accomplished by fluorescence-activated cell sorting.

7 . The method of claim 1 , wherein said cell population is expanded after said isolating.

8 . A method of producing a stem cell line, comprising transforming a stem cell with a DNA sequence that encodes a growth-promoting protein; and exposing said stem cell to conditions that promote production of said growth-promoting protein.

9 . The method of claim 8 , wherein said growth-promoting protein is v-myc, N-myc, c-myc, p53, SV40 large T antigen, polyoma large T antigen, E1a adenovirus or human papillomavirus E7 protein.

10 . The method of claim 8 , wherein said DNA sequence is regulatable.

11 . The method of claim 8 , wherein said DNA sequence is regulatable by tetracycline.

12 . The method of claim 8 , wherein said growth-promoting protein has a regulatable activity.

13 . The method of claim 8 , wherein said growth-promoting protein is a temperature-sensitive mutant.

14 . A method of producing a stem cell population, comprising identifying adherent placental stem cells that express one or more genes, wherein said genes are ACTG2, ADARB1, AMIGO2, ATRS-1, B4GALT6, BCHE, C11orf9, CD200, COL4A1, COL4A2, CPA4, DMD, DSC3, DSG2, ELOVL2, F2RL1, FLJ10781, GATA6, GPR126, GPRC5B, ICAM1, IER3, IGFBP7, IL1A, IL6, IL18, KRT18, KRT8, LIPG, LRAP, MATN2, MEST, NFE2L3, NUAK1, PCDH7, PDLIM3, PJP2, RTN1, SERPINB9, ST3GAL6, ST6GALNAC5, SLC12A8, TCF21, TGFB2, VTN, and ZC3H12A, at a detectably higher level than a bone marrow-derived mesenchymal stem cell that has undergone the same number of passages in culture as said placental stem cell, and isolating said placental cells.

15 . The method of claim 14 , wherein said adherent placental cell expresses five or more of said genes at a detectably higher level than a bone marrow-derived mesenchymal stem cell that has undergone the same number of passages in culture as said placental stem cell.

16 . The method of claim 14 , wherein said adherent placental cell expresses ten or more of said genes at a detectably higher level than a bone marrow-derived mesenchymal stem cell that has undergone the same number of passages in culture as said placental stem cell.

17 . The method of claim 14 , wherein said adherent placental cell expresses twenty or more of said genes at a detectably higher level than a bone marrow-derived mesenchymal stem cell that has undergone the same number of passages in culture as said placental stem cell.

18 . The method of claim 14 , wherein said adherent placental cell expresses each of said genes at a detectably higher level than a bone marrow-derived mesenchymal stem cell that has undergone the same number of passages in culture as said placental stem cell.

19 . A method of making a placental stem cell bank, comprising:

expanding primary culture placental stem cells from a human postpartum placenta for a first plurality of population doublings;

cryopreserving said placental stem cells to form a Master Cell Bank;

expanding a plurality of placental stem cells from the Master Cell Bank for a second plurality of population doublings;

cryopreserving said placental stem cells to form a Working Cell Bank;

expanding a plurality of placental stem cells from the Working Cell Bank for a third plurality of population doublings; and

cryopreserving said placental stem cells in individual doses,

wherein said individual doses collectively compose a placental stem cell bank.

20 . The method of claim 19 , wherein the total number of population doublings is about 20.

21 . The method of claim 19 , wherein said first plurality of population doublings is about four population doublings; said second plurality of population doublings is about eight population doublings; and said third plurality of population doublings is about eight population doublings.

22 . The method of claim 19 , wherein said primary culture placental stem cells comprise placental stem cells from placental perfusate.

23 . The method of claim 19 , wherein said primary culture placental stem cells comprise placental stem cells from digested placental tissue.

24 . The method of claim 19 , wherein said primary culture placental stem cells comprise placental stem cells from placental perfusate and from digested placental tissue.

25 . The method of claim 19 , wherein all of said placental stem cells in said placental stem cell primary culture are from the same placenta.

26 . The method of claim 19 , further comprising the step of selecting CD200 + or HLA-G + placental stem cells from said plurality of said placental stem cells from said Working Cell Bank to form individual doses.

27 . The method of claim 19 , wherein said individual doses comprise from about 10 4 to about 10 5 placental stem cells.

28 . The method of claim 19 , wherein said individual doses comprise from about 10 5 to about 10 6 placental stem cells.

29 . The method of claim 19 , wherein said individual doses comprise from about 10 6 to about 10 7 placental stem cells.

30 . The method of claim 19 , wherein said individual doses comprise from about 10 7 to about 10 8 placental stem cells.

31 . A composition comprising conditioned medium, wherein said medium is conditioned by placental stem cells that are:

CD200 + and HLA-G + ;

CD73 + , CD105 + , and CD200 + ;

CD200 + and OCT-4 + ;

CD73 + , CD105 + and HLA-G + ;

CD73 + and CD105 + and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow the formation of an embryoid-like body; or

OCT-4 + and facilitates the formation of one or more embryoid-like bodies in a population of placental cells comprising the stem cell when said population is cultured under conditions that allow formation of embryoid-like bodies;

or any combination of the foregoing.

32 . The composition of claim 31 , wherein said placental stem cells have been grown in said medium for at least one day.

33 . The composition of claim 31 , wherein said placental stem cells have been grown in said medium for at least three days.

34 . A composition comprising a placental stem cell that is:

CD200 + and HLA-G + ;

CD73 + , CD105 + , and CD200 + ;

CD200 + and OCT-4 + ;

CD73 + , CD105 + and HLA-G + ;

CD73 + and CD105+ and facilitates the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow the formation of an embryoid-like body; or

OCT-4 + and facilitates the formation of one or more embryoid-like bodies in a population of placental cells comprising the stem cell when said population is cultured under conditions that allow formation of embryoid-like bodies; or any combination of the foregoing; and

a stem cell that is not obtained from a placenta.

35 . The composition of claim 34 , wherein said stem cell not obtained from a placenta is an embryonic stem cell.

36 . The composition of claim 34 , wherein said stem cell not obtained from a placenta is a mesenchymal stem cell.

37 . The composition of claim 34 , wherein said stem cell not obtained from a placenta is a bone marrow-derived stem cell.

38 . The composition of claim 34 , wherein said stem cell not obtained from a placenta is a hematopoietic progenitor cell.

39 . The composition of claim 34 , wherein said stem cell not obtained from a placenta is a somatic stem cell.

40 . The composition of claim 39 , wherein said somatic stem cell is a neural stem cell, a hepatic stem cell, a pancreatic stem cell, an endothelial stem cell, a cardiac stem cell, or a muscle stem cell.

41 . A method of isolating a population of placental stem cells, comprising culturing a portion of a placenta for a time sufficient for a plurality of placental stem cells to proliferate out of said tissue, and isolating said placental stem cells from said tissue.

42 . The method of claim 41 , wherein said tissue is amniotic membrane, chorion, a combination of amnion and chorion, or a portion of any thereof, or a combination of any of the foregoing.

43 . An isolated adherent placental stem cell, wherein said stem cell is:

CD200 + and HLA-G + ;

CD73 + , CD105 + , and CD200 + ;

CD200 + and OCT-4 + ;

CD73 + , CD105 + and HLA-G + ;

CD73 + and CD105 + and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow the formation of an embryoid-like body; or

OCT-4 + and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising the stem cell when said population is cultured under conditions that allow formation of embryoid-like bodies, or a combination thereof;

and wherein said placental stem cell is non-maternal in origin.

44 . A population of isolated adherent placental stem cells, wherein said stem cells are:

CD200 + and HLA-G + ;

CD73 + , CD105 + , and CD200 + ;

CD200 + and OCT-4 + ;

CD73 + , CD105 + and HLA-G + ;

CD73 + and CD105 + and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow the formation of an embryoid-like body; or

OCT-4 + and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising the stem cell when said population is cultured under conditions that allow formation of embryoid-like bodies;

and wherein at least 70% of said placental stem cells are non-maternal in origin.

45 . The population of claim 44 , wherein at least 90% of said placental stem cells are non-maternal in origin.

46 . The population of claim 44 , wherein at least 99% of said placental stem cells are non-maternal in origin.

47 . A composition comprising the isolated placental stem cell of claim 43 .

48 . A composition comprising the isolated placental stem cell population of claim 44 .

49 . The composition of claim 47 that is in a form suitable for intravenous administration.

50 . The composition of claim 48 that is in a form suitable for intravenous administration.

51 . The composition of claim 47 wherein said composition is in a container suitable for intravenous administration of said composition.

52 . The composition of claim 48 wherein said composition is in a container suitable for intravenous administration of said composition.

53 . A method of collecting placental stem cells, comprising:

perfusing a mammalian placenta that has been drained of cord blood and perfused to remove residual blood;

perfusing said placenta with a perfusion solution through placental vasculature;

collecting said perfusion solution only from said vasculature, wherein said perfusion solution after perfusion comprises a population of cells that comprises placental stem cells, wherein at least 95% of said placental stem cells are fetal in origin;

and isolating a plurality of said placental stem cells from said population of cells.

54 . The method of claim 53 , wherein the perfusion solution is passed through the umbilical vein and collected from the umbilical arteries.

55 . The method of claim 53 , wherein the perfusion solution is passed through the umbilical arteries and collected from the umbilical vein.

56 . An isolated population of adherent placental stem cells collected by the method of claim 53 , wherein at least 95% of said isolated population of placental stem cells is fetal in origin.

57 . The isolated population of claim 51 , wherein greater than 99% of said placental stem cells are fetal in origin.

58 . The isolated population of claim 51 , wherein the placental stem cells are at least 50% of said population of placental cells.

59 . The isolated population of claim 51 , wherein the placental stem cells are at least 95% of said population of placental cells.