Monophenotypic in vitro cell lines of megakaryocytic...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/08 (2006.01) C07K 14/47 (2006.01) C07K 14/495 (2006.01) C07K 14/50 (2006.01) C07K 14/755 (2006.01) C12N 5/02 (2006.01) C12N 5/06 (2006.01)

Patent

CA 2077755

2077755 9113973 PCTABS00007 Human monophenotypic cloned cell lines of megakaryocytic lineage are established in vitro through use of adherent stromal cells in long term human bone marrow culture. One cloned in vitro cell line, CHRF-288-11, exhibits a karyotype and markers characteristic of megakaryocytes and platelets. CHRF-288-11 cells express platelet peroxidase, platelet factor IV, platelet CA++-ATPase, gpIIbIIIa, factor VIII, and MY7, MY9 and HLA-Dr antigens. CHRF-288-11 cell lines do not express erythroid markers glycophorin A and hemoglobin, myeloid marker myeloperoxidase, nor markers specific for T- and B-cells. CHRF-288-11 cells also express basic FGF and TGF-beta properties. CHRF-288-11 cells exhibit a constant karyotype (50, XY) and displays a doubling time of approximately 33 hours in either 25 % horse or calf serum. Treatment of CHRF-288-11 cells with phorbol 12-myristate 13-acetate (PMA) induces multi-nucleation and hyperploidy with about 35 % of the cells exhibiting two or more nuclei per cell, and greater than 80 % of the cells enlarging in size. CHRF-288-11 cell line is deposited under the Budapest Treaty with American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852, under accession no. CRL 10107.

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