Phenotypic characterization of the hematopoietic stem cell

• US 5,840,580 A
• Filed: 05/14/1997
• Issued: 11/24/1998
• Est. Priority Date: 05/01/1990
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Loken, Michael R., Huang, Shiang, Terstappen, Leon W., Lund-Johansen, Fridtjof, Olweus, Johanna
• Primary Examiner(s): Stanton, Brian R.
• Application Number:
  US08/856,406
• Time in Patent Office: 559 Days
• Field of Search: 424/93.1, 435/325, 435/326, 435/372
• US Class Current: 435/342
• 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: 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 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

One or more population of cells enriched for human hematopoietic stem cells is disclosed. HSC in this population of cells are capable of limited self-renewal and are capable of differentiating into all elements of the hematopoietic system. This population of cells has the phenotype of CD34⁺ /CD38^- and more preferably CD34⁺ /CD38^- /HLA-DR⁺. Cells within this population have been found to express CD13, CD33 and CD71. Hematopoietic stem cells can be used in a number of therapies, including autologous transplantation and in gene therapy.

Claims

1. A population of human cells that lacks mature progenitor cells of the erythroid, lymphoid, and myeloid lineages and is enriched for hematopoietic stem cells wherein said stem cells are capable of limited self-renewal and differentiation into erythroid myeloid and lymphoid progenitors and mature cells, which population is comprised of cells having a CD34⁺ /CD38^- /HLA-DR⁺ phenotype, wherein cells having said phenotype comprise at least 95% of said population.

2. The population of cells of claim 1 wherein said human cells are also at least one or more of CD13⁺, CD33⁺ and CD71⁺.

3. The population of human cells of claim 1 wherein said cells are derived from fetal or adult tissue.

4. The population of human cells of claim 3 wherein said cells are derived from peripheral blood, cord blood, bone marrow, liver or spleen.

5. A population of human cells comprising hematopoietic stem cells wherein said human cells are of a phenotype CD34⁺ /CD38^- /HLA-DR⁺ /CD13⁺ /CD33⁺ /CD71⁺, wherein cells of the phenotype comprise at least 95% of said population.

6. A method of isolating a population of human cells that lacks mature and progenitor cells of the erythroid, lymphoid and myeloid lineages and enriched for hematopoietic stem cells comprising the steps of obtaining human tissue containing hematopoietic stem cells, selecting said hematopoietic cells based upon the expression of at least CD34, HLA-DR, and CD38 antigens, and separating the cells that express CD34 and HLA-DR but do not express CD38, wherein said CD34⁺ /HLA-DR⁺ /CD38^- cells comprise at least 95% of said population.

7. The method of claim 6 wherein the selection step is carried out with monoclonal antibodies.

8. The method of claim 7 wherein the monoclonal antibodies are fluorescently labelled and the selection and separation steps are carried out by means of flow cytometry.

9. The method of claim 6 wherein mature erythroid cells are removed as a preliminary selection step.