Human hematopoietic stem cell

• US 5,061,620 A
• Filed: 03/30/1990
• Issued: 10/29/1991
• Est. Priority Date: 03/30/1990
• Status: Expired due to Term

• Current Assignee: Novartis Ag
• Original Assignee: Systemax Incorporated
• Inventors: Baum, Charles M., Aihara, Yukoh, Tsukamoto, Ann, Weissman, Irving
• Primary Examiner(s): Weimar, Elizabeth C.
• Assistant Examiner(s): Wang, Gian
• Application Number:
  US07/502,616
• Time in Patent Office: 578 Days
• Field of Search: 435/240.2, 435/240.21, 435/29, 435/240.25, 435/7.21
• US Class Current: 435/7.21
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• 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 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2502/1394 Bone marrow stromal cells; whole marrow
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: G01N 33/56972 White blood cells

Abstract

Human hematopoietic stem cells are provided by separation of the stem cells from dedicated cells. The stem cells may than be maintained by regeneration in an appropriate growth medium. Means are provided for assaying for the stem cells as to their capability for producing members of each of the hematopoietic lineages.

Claims

1. A cellular composition comprising human hematopoietic stem cells with fewer than 5% of lineage committed cells, wherein said hematopoietic stem cells are characterized as Thy-1⁺, capable of self-regeneration in a coculture medium and differentiation to members of the lymphoid and myelomonocytic hematopoietic lineages.

2. A cellular composition having at least 80% of the cells characterized by being human, hematopoietic and being CD34⁺ 10^- 19^- 33^-.

3. A cellular composition according to claim 2, wherein said cells are further characterized by being Thy-1⁺.

4. A method for obtaining a cellular composition comprising human hematopoietic stem cells, wherein fewer than about 100 cells is capable of self regeneration in a coculture medium and differentiation to members of the lymphoid and myelomonocytic hematopoietic lineages, said method comprising:

separating a mixture of human hematopoietic stem and differentiated cells into a substantially homogeneous fraction comprising cells characterized by being human, hematopoietic and being CD34⁺ 10^- 19^- 33^-.

5. A method according to claim 4, wherein said cells are further characterized by being Thy-1⁺.

6. A method according to claim 4, wherein said separation comprises:

combining said mixture with monoclonal antibodies conjugated to fluorescent labels to CD34, 10, 19 and 33, wherein said antibodies to 34 have a different fluorescent label from the other antibodies; and

separating a cell fraction characterized by being CD34⁺ 10^- 19^- 33^- by means of said fluorescent labels.

7. A method according to claim 6, comprising the additional step of combining said CD34⁺ 10^- 19^- 33^- cell fraction with a fluorescent labeled monoclonal antibody to Thy-1;

and

isolating a Thy-1⁺ fraction.