Use of stem cell factor and soluble interleukin-6 receptor for the ex vivo expansion of hematopoietic multipotential cells

• US 5,861,315 A
• Filed: 10/16/1996
• Issued: 01/19/1999
• Est. Priority Date: 11/16/1994
• Status: Expired due to Fees

• Current Assignee: Amgen Inc., Tosoh Corporation
• Original Assignee: Amgen Inc., Tosoh Corporation
• Inventors: Nakahata, Tatsutoshi
• Primary Examiner(s): ACHUTAMURTHY, PONNATHAPURA N
• Application Number:
  US08/732,023
• Time in Patent Office: 825 Days
• Field of Search: 424/93.1, 424/93.7, 435/325, 435/347, 435/378, 435/384, 435/386, 514/2, 514/21
• US Class Current: 435/384
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/125 Stem cell factor [SCF], c-kit ligand [KL]
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 5/0641 Erythrocytes
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

Stem cell factor in combination with soluble interleukin-6 receptor, interleukin-6, for gp130 signaling, supports the proliferation, differentiation and terminal maturation of blood cells from normal human hematopoietic multipotential cells.

Claims

1. A method for the ex vivo expansion of multipotential hematopoietic cells, comprising culturing the cells in a medium comprising soluble interleukin-6 receptor, interleukin-6, and stem cell factor, said cells, interleukin-6 receptor, interleukin-6, and stem cell factor derived from a human species, said interleukin-r receptor, interleukin-6, and stem cell factor being present in amounts effective to produce a mature erythroid cell population in the absence of erythropoietin.

2. The method according to claim 1, wherein the cells to be expanded are obtained from cord blood, peripheral blood or bone marrow.

3. The method according to claim 1, wherein the multipotential hematopoietic cells are progenitor and/or stem cells obtained by CD34 selection.

4. The method according to claim 1, wherein the multipotential hematopoietic cells are stem cells functionally selected by the removal of proliferating cells.

5. The method according to claim 1, wherein said soluble interleukin-6 receptor is used at a concentration of at least 80 ng/ml to 1280 ng/ml.

6. The method according to claim 5, wherein said interleukin-6 is used at a concentration of at least 50 ng/ml to 100 ng/ml.

7. A kit, consisting of:

soluble interleukin-6 receptor, interleukin-6 and stem cell factor, said soluble interleukin-6 receptor, interleukin-6 and stem cell factor provided in individual containers or as a mixture in a single container in an amount effective for use in expansion of multipotential hematopoietic cells according to claim 1.

8. A method generating differentiated blood cell colonies by ex vivo expansion of multipotential hematopoietic cells, comprising culturing the hematopoietic cells in a medium comprising soluble interleukin-6 receptor, interleukin-6, and stem cell factor, said hematopoietic cells, interleukin-6 receptor, interleukin-6, and stem cell factor derived from a human species, said interleukin-6 recaptor, interleukin-6, and stem cell factor being present in amounts effective to produce a mature erythroid cell population in the absence of erythropoietin.