Placental stem cells

• US 8,580,563 B2
• Filed: 10/27/2008
• Issued: 11/12/2013
• Est. Priority Date: 12/06/2000
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Hariri, Robert J.
• Primary Examiner(s): LI, QIAN JANICE
• Application Number:
  US12/259,259
• Publication Number:
  US 20090053805A1
• Time in Patent Office: 1,842 Days
• Field of Search: None
• US Class Current: 435/366
• CPC Class Codes: A01N 1/021 Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61M 1/02 Blood transfusion apparatus blood infusion by syringes A61M5/14
• CPC Class Codes: A61M 2202/0462 Placental blood, umbilical cord blood
• CPC Class Codes: A61M 2210/1458 Placenta
• CPC Class Codes: A61M 2210/1466 Umbilical cord
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/00 Drugs for disorders of the urinary system
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 : Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 : Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/14 : of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12M 29/10 : Perfusion
• CPC Class Codes: C12M 47/04 : Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12N 2509/00 : Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2510/02 : Cells for production
• CPC Class Codes: C12N 2533/90 : Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0603 : Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0605 : Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: C12N 5/0668 : Mesenchymal stem cells from other natural sources

Abstract

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.

Claims

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.

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.

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.