Method of collecting placental stem cells

• US 7,045,148 B2
• Filed: 12/05/2001
• Issued: 05/16/2006
• Est. Priority Date: 12/06/2000
• Status: Expired due to Term

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Hariri, Robert J.
• Primary Examiner(s): LI, QIAN JANICE
• Application Number:
  US10/004,942
• Publication Number:
  US 20020123141A1
• Time in Patent Office: 1,623 Days
• Field of Search: 435/2, 435/325, 435/395, 435/41, 435/69.1, 435/70.1, 435/70.3, 435/1.1, 435/1.2
• US Class Current: 424/529
• 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 CD34⁺ stem cells from an isolated and exsanguinated mammalian placenta, said method comprising:

perfusing said placenta with a perfusion solution in an amount and for a time sufficient to collect a detectable amount of said cells comprising CD34⁺ stem cells, wherein said placenta has been drained of cord blood and flushed to remove residual blood prior to said perfusing, wherein said CD34⁺ stem cells are not obtained from cord blood, and wherein said perfusing is performed by passing said perfusion solution into one or both of the umbilical artery and umbilical vein of said placenta; and

collecting said CD34⁺ stem cells and perfusion solution from said placenta.

2. The method of claim 1, wherein said perfusing is performed at at least four hours after removal of said residual blood.

3. The method of claim 1, wherein said perfusing is performed at at least twelve hours after removal of said residual blood.

4. The method of claim 1, wherein said perfusing is performed at at least 24 hours after removal of said residual blood.

5. The method of claim 1, wherein said perfusing is performed using a first volume of between about 30 ml and about 150 ml of said perfusion solution.

6. The method of claim 5, further comprising continuing said perfusing using a second volume of about 30 ml to about 150 ml of said perfusion solution, said second volume being collected separately from said first volume.

7. The method of claim 1, in which said perfusing is performed for a plurality of times.

8. The method of claim 7, wherein, for each of said times, said perfusing is performed using a volume of about 30 ml to about 150 ml of said perfusion solution.

9. The method of claim 1, wherein said perfusion solution comprises an anticoagulant.

10. The method of claim 5, wherein said perfusion solution comprises an anticoagulant.

11. The method of claim 7, wherein said perfusion solution comprises an anticoagulant.

12. The method of claim 1, wherein said perfusion solution comprises heparin, ethylene diamine tetra acetic acid (EDTA) or creatine phosphate dextrose (CPDA).

13. The method of claim 1, wherein said perfusion solution comprises a growth factor or a cytokine.

14. The method of claim 13, wherein said growth factor or cytokine is selected from the group consisting of a colony stimulating factor, interferon, erythropoietin, stem cell factor, thrombopoietin, an interleukin, granulocyte colony-stimulating factor, and any combination thereof.

15. The method of claim 1, wherein said isolated mammalian placenta is a post-partum placenta remaining after a successful birth.

16. The method of claim 1, further comprising separating said CD34⁺ stem cells from cells other than CD34⁺ stem cells and said perfusion solution.

17. The method of claim 5, further comprising separating said CD34⁺ stem cells from said perfusion solution.

18. The method of claim 6, further comprising separating said CD34⁺ stem cells from said perfusion solution.

19. The method of claim 5, wherein said CD34⁺ stem cells are collected over a period of up to 48 hours.

20. The method of claim 6, wherein said CD34⁺ stem cells are collected over a period of up to 48 hours.