Hematopoietic cells: compositions and methods
AssigneeThe Regent of the University of Michigan
InventorsRussell S. Taichman, Stephen G. Emerson
Life of this patent
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Abstract
Processes, compositions and uses of hematopoietic cells are disclosed. Hematopoietic cells are cells which can differentiate into mature blood cells when co-cultured with osteoblasts. Specifically, a process for propagating and maintaining the immature morphology of a hematopoietic cell by co-culturing with osteoblasts is disclosed. The osteoblasts provide cytokines and/or a microenvironment which propagates and maintains the immature morphology of a hematopoietic cell. Hematopoietic cells are useful in the treatment of certain blood related disorders and are useful for treatment of patients in need of hematopoietic cells.
Claims41
A process for propagating and maintaining the immature morphology of mammalian hematopoietic cells, said immature morphology being defined as CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.-, the process comprising the steps of:
- (a) obtaining an enriched population of mammalian hematopoietic cells having the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- ;
- (b) co-culturing said enriched population of mammalian hematopoietic cells in the presence of osteoblast cells for a time period of between about 2 weeks and about 8 weeks;
- and
- (c) collecting mammalian hematopoietic cells having the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- maintained by the process of step (b).
The process of claim 26, wherein said enriched population of mammalian hematopoietic cells is co-cultured in the presence of osteoblast cells for a time period of between about 2 weeks and about 6 weeks.
A process for propagating mammalian hematopoietic cells that maintain an immature morphology defined as CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.-, the process comprising the steps of:
- (a) obtaining an enriched population of mammalian hematopoietic cells having the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- by a method comprising the enrichment processes of:
- (i) subjecting mammalian bone marrow cells to equilibrium-density centrifugation to provide centrifuged cells with a density of between about 1.010 and about 1.090 grams/cm.sup.3 ;
- and
- (ii) exposing the centrifuged cells to an anti-CD34 antibody immunoreactive with the hematopoietic cell antigen CD34, and removing cells that do not immunoreact with the anti-CD34 antibody;
- (b) co-culturing said enriched population of mammalian hematopoietic cells in the presence of osteoblast cells for a time period of between about 2 weeks and about 8 weeks;
- and
- (c) collecting mammalian hematopoietic cells having the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- maintained by the process of step (b).
A process for propagating mammalian hematopoietic cells that maintain the immature morphology defined as CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.-, the process comprising the steps of:
- (a) obtaining an enriched population of mammalian hematopoietic cells having the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- by a method comprising the enrichment processes of:
- (i) subjecting mammalian bone marrow cells to equilibrium-density centrifugation to provide centrifuged cells with a density of between about 1.040 and about 1.080 grams/cm.sup.3 ;
- (ii) exposing the centrifuged cells to an adherent surface, and removing adherent cells to provide centrifuged, non-adherent cells;
- and
- (iii) exposing the centrifuged, non-adherent cells to an anti-CD34 antibody immunoreactive with the hematopoietic cell antigen CD34, and removing cells that do not immunoreact with the anti-CD34 antibody;
- (b) co-culturing said enriched population of mammalian hematopoietic cells in the presence of osteoblast cells for a time period of between about 2 weeks and about 8 weeks;
- and
- (c) collecting mammalian hematopoietic cells having the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- maintained by the process of step (b).
A purified population of mammalian hematopoietic cells that maintain the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- for a time period of between about 2 weeks and about 8 weeks, said hematopoietic cells having a density of between about 1.010 gm/cm.sup.3 and about 1.090 gm/cm.sup.3 and being prepared by a process comprising the steps of:
- (a) obtaining an enriched population of mammalian hematopoietic cells having the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- ;
- (b) co-culturing said enriched population of mammalian hematopoietic cells in the presence of osteoblast cells for a time period of between about 2 weeks and about 8 weeks;
- and
- (c) obtaining therefrom a population of mammalian hematopoietic cells that have maintained the immature morphology of CD34.sup.+, HLA-DR.sup.+, Thy-1.sup.+ and Lin.sup.- and have a density of between about 1.010 gm/cm.sup.3 and about 1.090 gm/cm.sup.3.
Dependencies are inferred from the claim text.