Methods for augmentation collection of placental hematopoietic stem cells and uses thereof

• US 8,574,899 B2
• Filed: 12/12/2011
• Issued: 11/05/2013
• Est. Priority Date: 12/22/2010
• Status: Active Grant

• Current Assignee: Frans A. Kuypers, Vladimir B. Serikov
• Original Assignee: Frans A. Kuypers, Vladimir B. Serikov
• Inventors: Serikov, Vladimir B, Kuypers, Frans A
• Primary Examiner(s): Lankford, Blaine
• Assistant Examiner(s): Visone, Thomas J
• Application Number:
  US13/374,105
• Publication Number:
  US 20120177616A1
• Time in Patent Office: 694 Days
• Field of Search: None
• US Class Current: 435/325
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• 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 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The subject matter of this invention is a novel method to augment the process of obtaining populations of placental hematopoietic stem and progenitor cells for use in medical practices. A method of augmentation of stem cells collection from placenta is claimed comprising the steps of (a) infusing placental vessels cell preservation compound and a with a composition containing blockers of cell adhesion receptors, (b) incubating said placenta for a sufficient period of time, (c) placing placenta in a containment with sufficient intensity of electromagnetic or ultrasound field for a sufficient period of time; (d) eluting cell suspension from placental vessels, (e) collecting cell suspension and harvesting cells. Invention further claims a method of treatment a disease by means of administering therapeutic composition containing said placental-derived hematopoietic stem cells.

Claims

1. A method of obtaining hematopoietic stem cells from a placenta, comprising the steps of:

a. infusing placental tissue with a composition comprising a cell preservation compound to provide infused placental tissue without simultaneous effusion;

b. incubating said infused placental tissue for a duration to provide incubated placental tissue that releases hematopoietic stem cells, wherein said duration is selected from the group consisting of;

i. about 2 hours to about 3 hours;

ii. about 3 hours to about 4 hours;

iii. about 4 hours to about 6 hours;

iv. about 6 hours to about 8 hours; and

v. about 8 hours to about 12 hours; and

c. isolating hematopoietic stems cells from the incubated placental tissue;

wherein the infused placental tissue or incubated placental tissue is exposed to at least one of ultrasonic radiation or an electromagnetic field.

2. The method of claim 1, wherein the method comprises exposing the infused placental tissue or incubated placental tissue to ultrasonic radiation.

3. The method of claim 1, wherein the method comprises exposing the infused placental tissue or incubated placental tissue to an electromagnetic field.

4. The method of claim 3, wherein the electromagnetic field has a strength of at least 0.5 gauss.

5. The method of claim 1, wherein said incubating is performed at a temperature of about 20°

C. for at least 2 hours.

6. The method of claim 1, wherein said isolating comprises (i) eluting a suspension of cells from the incubated placental tissue to provide a cellular suspension, and (ii) subjecting the cellular suspension to centrifugal separation and thereafter harvesting hematopoietic stem cells.

7. The method of claim 1, wherein said composition further comprises at least one cell adhesion receptor blocker.

8. The method of claim 1, wherein the cell preservation compound is an antioxidant.

9. The method of claim 7, wherein the cell adhesion receptor blocker is a CXCR4 receptor antagonist, an integrin inhibitor, a biocompatible non-ionic detergent, a PCAM receptor antagonist, a PCAM receptor antibody, an ICAM-blocking compound, a VCAM-blocking compound, a CD56 (NCAM)-blocking compound, a proteolytic enzyme, a fibronectin receptor-blocking compound, or a calcium chelator.

10. The method of claim 7, wherein the cell adhesion receptor blocker is etoposide.

11. The method of claim 1, wherein the hematopoietic stem cells have phenotype CD34⁺CD45^dim.

13. The method of claim 1, wherein said incubating is performed for a period of time selected from the group consisting of:

a. about 4 hours to about 6 hours;

b. about 6 hours to about 8 hours; and

c. about 8 hours to about 12 hours.

14. The method of claim 1, wherein said incubating is performed for a period of time of about 8 hours to about 12 hours.

12. A method of obtaining hematopoietic stem cells from a placenta, comprising the steps of:

a. infusing a placenta or placental tissue with a cell-liberating composition comprising a cell preservation compound without simultaneous effusion, and then intermittently exposing the placenta or placental tissue to ultrasonic radiation while incubating the placenta or placental tissue for a duration ranging from 3 hours to 8 hours to provide incubated placental tissue that releases hematopoietic stem cells; and

b. isolating hematopoietic stems cells from the incubated placental tissue.