Method of collecting placental stem cells

• US 20020123141A1
• Filed: 12/05/2001
• Published: 09/05/2002
• 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.
• Granted Patent: US 7,045,148 B2
• Time in Patent Office: Days
• 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

A method of collecting embryonic-like stem cells from a placenta which has been treated to remove residual cord blood by perfusing the drained placenta with an anticoagulant solution to flush out residual cells, collecting the residual cells and perfusion liquid from the drained placenta, and separating the embryonic-like cells from the residual cells and perfusion liquid. Exogenous cells can be propagated in the placental bioreactor and bioactive molecules collected therefrom.

Claims

1. A method of collecting embryonic-like stem cells from a placenta which has been treated to remove residual cord blood said method comprising:

perfusing a placenta which has been drained of cord blood with an anticoagulant perfusion solution to flush out residual cells and embryonic-like stem cells from said drained placenta, and

collecting said residual cells and embryonic-like stem cells and perfusion solution from the drained placenta.

2. The method of claim 1 further comprising separating said embryonic-like stem cells from said residual cells and anticoagulant perfusion solution.

3. The method of claim 1 wherein the placenta is perfused with the anticoagulant solution by passing the anticoagulant solution into one or both of the umbilical artery and umbilical vein of said placenta.

4. The method of claim 1 including collecting said residual cells and said anticoagulant perfusion solution from the part of the placenta that was attached to the wall of the uterus of the mother.

5. The method of claim 2 including said embryonic-like stem cells are separated from said residual cells and anticoagulant perfusion solution by centrifugation.

6. The method of claim 1 including

continuing perfusion of said placenta for maintaining placental viability,

flushing out additional residual cells and additional embryonic-like stem cells,

collecting said additional residual cells, said additional embryonic-like stem cells and said additional anticoagulant perfusion solution, and

separating said additional embryonic-like stem cells from said additional residual cells.

7. The method of claim 6 including maintaining said perfusion of said placenta for stimulating the production of new cells and bioactive molecules, and collecting said newly produced cells and said bioactive molecules from said placenta.

8. The method of claim 6 wherein the residual cells and embryonic-like stem cells are collected over a period of 24 to 48 hours after birth.

9. The method of claim 6 wherein the placenta or cells in the placenta are stimulated to produce bioactive molecules.

10. The method of claim 7 wherein said newly produced cells and bioactive molecules are collected over a period of 24 to 48 hours after birth.

11. The method of claim 6 wherein the cells in the placenta are genetically engineered with exogenous DNA.

12. The method of claim 7 wherein the newly produced cells are genetically engineered with exogenous DNA.

14. The method of claim 13 wherein the remaining viable endogenous cells are removed by irradiation of the perfused placenta.

15. The method of claim 14 wherein the irradiation is selected from electromagnetic, ultra-violet, X-ray, gamma-, and beta-radiation.

16. The method of claim 13 wherein the exogenous cells are stem cells.

17. The method of claim 16 wherein the stems cells are hemopoietic pluripotent stem cells.

18. The method of claim 13 wherein the exogenous cells are stimulated to produce bioactive molecules.

19. The method of claim 18 wherein the bioactive molecules are collected prior to collecting the propagated exogenous cells.

20. The method of claim 13 wherein the exogenous cells are stimulated to propagate by introducing growth factors selected from cytokines, hormones, promoters, repressors, proteins, nucleic acids, viruses, and immunogens.

21. The method of claim 13 wherein the exogenous cells are selected from animal cells, insect cells, or human cells.

22. The method of claim 13 wherein the exogenous cells in the placenta are genetically engineered with exogenous DNA prior to collecting or during propagation in the perfused placental bioreactor.

23. The method of claim 22 wherein the exogenous DNA is introduced into the exogenous cells propagated in the perfused placenta via a vector selected from viral, adenoviral, and retroviral vectors.

24. The method of claim 22 wherein the exogenous DNA is introduced into the exogenous cells by a method selected from mechanical introduction of the exogenous DNA, chemical mediated uptake of the exogenous DNA, and liposomal mediated uptake of the exogenous DNA.

13. A method of propagating exogenous cells in a placental bioreactor which has been treated to remove residual umbilical cord blood, comprising:

perfusing a placenta which has been drained of cord blood with an anticoagulant perfusion solution to flush out residual cells and embryonic-like stem cells,

collecting said residual cells, said embryonic-like stem cells, and said anticoagulant perfusion solution from the drained placenta,

removing all remaining viable endogenous cells from the drained placenta,

perfusing the placenta with nutrient perfusion solution,

introducing exogenous cells into the perfused placenta,

incubating the perfused placenta with exogenous cells, and

collecting propagated exogenous cells from the perfused placenta.