Method of preparing an undifferentiated cell

• US 9,068,164 B1
• Filed: 05/10/2000
• Issued: 06/30/2015
• Est. Priority Date: 02/02/1995
• Status: Expired due to Fees

• Current Assignee: Tristem Trading (Cyprus) Limited
• Original Assignee: Tristem Trading (Cyprus) Limited
• Inventors: Abuljadayel, Ilham Saleh
• Primary Examiner(s): CANELLA, KAREN A
• Application Number:
  US09/568,254
• Time in Patent Office: 5,529 Days
• Field of Search: 435/325, 435/347, 435/355, 435/366, 435/372, 435/372.1, 435/372.2, 435/372.3, 435/374, 435/375, 435/377, 435/383, 435/384, 530/387.1, 530/388.1, 530/388.2, 5303887-38873, 530/388.75, 530/388.85, 424/130.1, 424/85.1
• US Class Current: 1/1
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood 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: C12N 2500/14 Calcium; Ca chelators; Calcitonin
• CPC Class Codes: C12N 2500/24 Iron; Fe chelators; Transferrin
• CPC Class Codes: C12N 2501/06 Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide
• CPC Class Codes: C12N 2501/14 Erythropoietin [EPO]
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/50 Cell markers; Cell surface determinants
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• CPC Class Codes: C12N 5/0635 B lymphocytes
• CPC Class Codes: C12N 5/0636 T lymphocytes
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

Disclosed is a method of preparing an undifferentiated cell. The method includes contacting a more committed cell with an agent that causes the more committed cell to retrodifferentiate into an undifferentiated cell.

Claims

1. A method comprising increasing the relative number of hematopoietic stem cells in a cell population including committed cells, wherein the method comprises contacting in vitro a cell population derived from unmobilized blood with an antibody that binds to MHC class II antigens, and enriching hematopoietic stem cells or recovering the hematopoietic stem cells from the cell population by using a cell surface marker, wherein the hematopoietic stem cells are MHC class I⁺ and/or MHC class II⁺ cells and wherein the hematopoietic stem cells are enriched or recovered from the cell population 2 hours after the cell population is first contacted with the antibody.

2. The method according to claim 1 wherein the committed cells are noncancer cells.

3. The method according to claim 1 wherein the committed cells are differentiated cells.

4. The method according to claim 3 wherein the committed cells are selected from CFC-T cells, CFC-B cells, CFC-Eosin cells, CFC-Bas cells, CFC-GM cells, CFC-MEG cells, BFC-E cells, CFC-E cells, T cells and B cells.

5. A method according to claim 1 wherein the antibody is a monoclonal antibody.

6. A method according to claim 5 wherein the antibody is selected from the group consisting of monoclonal antibody CR3/43 and monoclonal antibody TAL 1B5.

7. A method according to claim 1, wherein the antibody is used in conjunction with a biological response modifier, wherein the biological response modifier is selected from the group consisting of an alkylating agent, an immunomodulator, a growth factor, a cytokine, and a hormone.

8. A method comprising increasing the relative number of hematopoietic stem cells in a cell population including committed cells, wherein the method comprises contacting in vitro a cell population derived from unmobilized blood with an antibody that binds to MHC class II antigens, in conjunction with cyclophosphamide, wherein the hematopoietic stem cells are MHC class I⁺ and/or MHC class II⁺ cells.

9. A method comprising increasing the relative number of hematopoietic stem cells in a cell population derived from unmobilized blood, which population includes committed cells, said hematopoietic stem cells being capable of being committed into more differentiated hematopoietic cells, which method comprises:

(i) contacting in vivo and initial cell population comprising the committed cells with an antibody that binds to MHC class II antigens,

(ii) culturing the cell population thereby producing an altered cell population comprising hematopoietic stem cells, and

(iii) enriching said hematopoietic stem cells or recovering said hematopoietic stem cells from the altered cell population, wherein said hematopoietic stem cells are enriched or recovered from the altered cell population 2 hours after commencement of the incubation.

10. The method according to claim 9 wherein step (iii) comprises enriching said hematopoietic stem cells or recovering said hematopoietic stem cells from the altered cell population by using a cell surface marker.

11. A method comprising increasing the relative number of hematopoietic stem cells in a cell population including committed cells, wherein the method comprises contacting in vitro a cell population derived from unmobilized blood with an agent selected from the group consisting of (a) erythropoietin in combination with an antibody that binds to MHC class II antigens, and (b) GM-CSF in combination with an antibody that binds to MHC class II antigens, and enriching hematopoietic stem cells or recovering the hematopoietic stem cells from the cell population by using a cell surface marker,

wherein the hematopoietic stem cells are MHC class I⁺ and/or MHC class II⁺ cells, and

wherein the hematopoietic stem cells are enriched or recovered from the cell population 2 hours after the cell population is first contacted with the agent.

12. The method according to claim 11 wherein the committed cells are noncancer cells.

13. The method according to claim 11 wherein the committed cells are differentiated cells.

14. The method according to claim 13, wherein the committed cells are selected from CFC-T cells, CFC-B cells, CFC-Eosin cells, CFC-Bas cells, CFC-GM cells, CFC-MEG cells, BFC-E cells, CFC-E cells, T cells and B cells.

15. The method according to claim 11 wherein the antibody is a monoclonal antibody.

16. The method according to claim 15 wherein the antibody is selected from the group consisting of monoclonal antibody CR3/43 and monoclonal antibody TAL 1B5.

17. A method according to claim 11, wherein the agent is used in conjunction with a biological response modifier, wherein the biological response modifier is selected from the group consisting of an alkylating agent, an immunomodulator, a growth factor, a cytokine, and a hormone.

20. The method of claim 11 or 18, wherein the agent comprises erythropoietin in combination with an antibody that binds to MHC class II antigens.

21. The method of claim 20, wherein the antibody that binds to MHC class II antigens is an antibody that binds to the homologous region of the beta chain of MHC class II antigens.

22. The method of claim 11 or 18, wherein the agent comprises GM-CSF in combination with an antibody that binds to MHC class II antigens.

23. The method of claim 22, wherein the antibody that binds to MHC class II antigens is an antibody that binds to the homologous region of the beta chain of MHC class II antigens.

18. A method comprising increasing the relative number of hematopoietic stem cells in a cell population derived from unmobilized blood, which population includes committed cells, said hematopoietic stem cells being capable of being committed into more differentiated hematopoietic cells, which method comprises:

(i) contacting in vivo and initial cell population comprising the committed cells with an agent selected from the group consisting of (a) erythropoietin in combination with an antibody that binds to MHC class II antigens, and (b) GM-CSF in combination with an antibody that binds to MHC class II antigens,

(ii) culturing the cell population thereby producing an altered cell population comprising hematopoietic stem cells, and

(iii) enriching said hematopoietic stem cells or recovering said hematopoietic stem cells from the altered cell population, wherein said hematopoietic stem cells are enriched or recovered from the altered cell population 2 hours after commencement of the incubation.

19. The method according to claim 18 wherein step (iii) comprises enriching said hematopoietic stem cells or recovering said hematopoietic stem cells from the altered cell population using a cell surface marker.

24. A method comprising increasing the relative number of hematopoietic stem cells in a cell population including committed cells, wherein the method comprises contacting in vitro a cell population derived from unmobilized blood with an agent selected from the group consisting of (a) erythropoietin in combination with an antibody that binds to MHC class II antigens, and (b) GM-CSF in combination with an antibody that binds to MHC class II antigens, in conjunction with cyclophosphamide, wherein the hematopoietic stem cells are MHC class I⁺ and/or MHC class II⁺ cells.