Procurement, isolation and cryopreservation of maternal placental cells

• US 20080064098A1
• Filed: 06/05/2007
• Published: 03/13/2008
• Est. Priority Date: 06/05/2006
• Status: Abandoned Application

• Current Assignee: Cryo-Cell International Incorporated
• Original Assignee: Cryo-Cell International Incorporated
• Inventors: Allickson, Julie
• Application Number:
  US11/810,932
• Publication Number:
  US 20080064098A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly

Abstract

Methods, processes and systems for procuring, isolating and cryopreserving at least one viable, multipotent maternal placental stem cell is provided. Viable maternal placental stem cells are also provided. The maternal placental stem cell of the invention expresses the cell surface marker CD117 and at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD166, SSEA-3 and SSEA-4 and has low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45, CD133, TRA-1-60 and TRA-1-81. The methods and process comprise generally obtaining a piece of placental tissue from a whole placenta, disaggregating the placental tissue with mechanical separation or enzymatic digestion, collecting and concentrating placental cells comprising maternal placental cells with centrifugation and optionally cryopreserving the placental cells. Additional steps for selecting and culturing the maternal placental stem cells is provided.

Claims

1. A method for obtaining a population of cells comprising maternal placental stem cells, the method comprising:

(a) disaggregating placental tissue and separating the population of cells from disaggregated placental tissue;

(b) collecting and concentrating the population of cells; and

(c) cryopreserving the population of placental cells at or below about −

135°

.

2. The method of claim 1, wherein the maternal placental stem cells express the cell surface marker CD117 and at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD166, SSEA-3 and SSEA-4 and have low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45 and CD133 from placental tissue.

3. A process for obtaining a population of cells comprising maternal placental stem cells, the method comprising:

(a) procuring placental tissue from a whole placenta, the placental tissue comprising maternal tissue and fetal tissue;

(b) disaggregating the placental tissue;

(c) isolating the population of cells comprising maternal placental cells from disaggregated maternal tissue; and

(d) collecting the population of cells comprising maternal placental stem cells by concentrating the population of placental cells with at least one step of centrifugation.

4. The process of claim 3, wherein the maternal placental stem cells express the cell surface marker CD117 and at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD166, SSEA-3 and SSEA-4 and have low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45 and CD133 from placental tissue.

5. A process for collecting maternal placental stem cells expressing CD117 from placental tissue, the process comprising:

(a) isolating placental cells comprising the maternal placental stem cells from the placental tissue by disaggregating the placental tissue;

(b) collecting and concentrating the placental cells; and

(c) cryopreserving the placental cells.

6. The process of claim 5, wherein the maternal placental stem cells also express at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD166, SSEA-3 and SSEA-4 and have low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45 and CD133 from placental tissue.

7. A system for collecting a population of cells comprising maternal placental stem cells, comprising:

(a) a placental cell isolater, wherein the placental cell isolator disaggregates placental tissue comprising maternal tissue and separates placental cells from the disaggregate placental tissue;

(b) a placental cell collector, wherein the placental cell collector collects the placental cells separated from the disaggregate placental tissue;

(c) a placental cell concentrator, wherein the placental cell concentrator concentrates placental cells present in a suspension; and

(d) a placental cell cryopreserver, wherein the placental cell cryopreserver maintains the collected and concentrated placental cells at a temperature at or below about −

135°

C.

8. A process for isolating maternal placental stem cells expressing CD117 from a population of placental cells, the process comprising:

(a) culturing a population of placental cells comprising maternal placental stem cells;

(b) selecting placental cells expressing CD117 from a culture of the population of placental cells; and

(c) cryopreserving the maternal placental stem cells expressing CD117.

9. The process of claim 8, wherein the maternal placental stem cells also express at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD166, SSEA-3 and SSEA-4 and have low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45 and CD133 from placental tissue.

10. A process for isolating a population of maternal placental stem cells from a population of placental cells, comprising, selecting maternal placental stem cells expressing CD117 from a culture of the population of placental cells.

11. A process for isolating a population of maternal placental stem cells expressing CD117 from a population of placental cells, the process comprising,

(a) selecting placental cells expressing CD117 from a population of placental cells comprising maternal placental stem cells;

(b) culturing the placental cells expressing CD117 selected from the population of placental cells comprising maternal placental stem cells; and

(c) selecting placental cells expressing CD117 from a culture of placental cells.

12. A population of cells enriched for maternal placental stem cells obtained from the process comprising:

(a) culturing a population of cells comprising maternal placental stem cells;

and

(b) selecting cells expressing CD117 from a culture of the population of cells.

13. A population of maternal placental stem cells obtained from the process comprising selecting placental cells expressing CD117 from a culture of a population of placental cells.

14. The population of maternal placental cells of claim 13, wherein the maternal placental stem cells express the cell surface marker CD117 and at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD166, SSEA-3 and SSEA-4 and have low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45 and CD133 from placental tissue.

15. Maternal placental stem cells obtained from the process comprising

(a) selecting placental cells expressing CD117 from a population of placental cells comprising maternal placental stem cells;

(b) culturing the placental cells expressing CD117 selected from the population of placental cells comprising maternal placental stem cells; and

(c) selecting placental cells expressing CD117 from a culture of placental cells expressing CD117.

16. A population of cells enriched for maternal placental stem cells expressing CD117.

17. A population of cells enriched for maternal placental stem cells express the cell surface marker CD117 and at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD166, SSEA-3 and SSEA-4 and have low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45 and CD133 from placental tissue.

18. A population of cells enriched for maternal placental stem cells expressing CD44 and CD117 and having low or no expression of CD45.

19. A composition comprising a population of cells enriched for maternal placental stem cells and a preservation agent.

20. A composition comprising at least one maternal placental stem cell and a preservation agent.

21. The composition of claim 20, wherein the at least one maternal placental stem cell expresses at least one of the cell surface markers selected from the group consisting of CD29, CD44, CD73, CD90, CD105, CD117, CD166, SSEA-3 and SSEA-4 and has low or no expression of at least one of the cell surface markers selected from the group consisting of CD34, CD45 and CD133 and a preservation agent.

22. At least one maternal placental stem cell obtained from the process comprising:

(a) procuring placental tissue comprising maternal tissue and fetal tissue from a whole placenta;

(b) disaggregating the placental tissue;

(c) isolating placental cells comprising maternal placental stem cells from disaggregated placental tissue;

(d) collecting and concentrating placental cells comprising maternal placental stem cell in a population of cells;

(e) culturing the population of placental cells comprising maternal placental stem cell; and

(f) selecting at least one maternal placental stem cell expressing CD117 from a culture of the population of placental cells.

23. The process of claim 22, wherein the process comprises the further steps of:

(a) culturing at least one maternal placental stem cell expressing CD117 selected from the population of placental cells; and

(b) selecting at least one maternal placental stem cell expressing CD117 from a culture of placental cells expressing CD117 comprising a population of at least one maternal placental stem cell expressing CD117.