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US5198356A Expired / Lapsed US ⏱️ 0.0 years remaining

Monophenotypic in vitro cell lines of megakaryocytic lineage, products produced thereby and methods

Assignee: CINCINNATI A CORP OF OH, University of
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Filing: 1990-03-12
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Granted: 1993-03-30
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Expires: 2010-03-12
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Abstract

Abstract 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. Long term bone marrow cultures are used for initial adaptation of the cells to culture conditions. Once adapted, cells of the human in vitro cell lines are weaned from the stromal layer until they proliferate in the complete absence of any feeder layer. Seed cells for establishment of human in vitro cell lines were derived from a human solid tumor designated as ATCC CRL 9139 xenograft. Cells of one cloned in vitro cell line designated as CHRF-288-11 exhibits a karyotype and markers characteristic of megakaryocytes and platelets. Cells of the CHRF-288-11 cell line express platelet peroxidate, platelet factor IV, platelet Ca++ -ATPase, gpIIbIIIa, factor VIII, and MY7, MY9 and HLA-Dr antigens. CHRF-288-11 cell line exhibits a constant karyotype (50, XY). CHRF-288-11 cell line is deposited under the Budapest Treaty with American Type Culture Collection, 12301 Parklawn Drive, Rockville, Md. 20852, under accession No. CRL 10107.

Inventors

  • 👤 Michael A. Lieberman
  • 👤 Douglas A. Fugman

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-23 15:18
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0634; C12N5/0644; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/495; C07K14/50; C07K14/503; C12N2502/00; C12N2502/13; C12N2502/1394

Claims Analysis

Total Claims: 11
Claim 1
Independent Claim 81 words
A continuous human in vitro cell line derived from cells designated ATCC CRL 9139 comprising monophenotypic cells of megakaryoctic lineage and having a constant karyotype of 50 XY having characteristics similar to colony forming unit megakaryoblasts, and approximately 80% of said cells have the ability to express platelet peroxidase, said cells further having at least one of the following characteristics: a. granulated cytoplasm, said granules being characteristic for alpha-granules; b. expresses gpIIbIIIa; c. expresses Factor VIII; and d. platelet Factor IV.
Claim 2
Dependent Claim 81 words
A continuous human in vitro cell line derived from cells designated ATCC CRL 9139 comprising monophenotypic cells of megakaryoctic lineage and having a constant karyotype of 50 XY having characteristics similar to colony forming unit megakaryoblasts, and approximately 80% of said cells have the ability to express platelet peroxidase, said cells further having at least one of the following characteristics: a. granulated cytoplasm, said granules being characteristic for alpha-granules; b. expresses gpIIbIIIa; c. expresses Factor VIII; and d. platelet Factor IV.
Claim 3
Dependent Claim 22 words
A human in vitro cell line of claim 1, said cells having a diameter in the range of between about 15-20 microns.
Claim 4
Dependent Claim 17 words
A human in vitro cell line of claim 1, said cells being derived from human extramedullary tissue.
Claim 5
Dependent Claim 37 words
A human in vitro cell line of claim 1, certain of said cells having the ability to express at least one protein selected from the group consisting of basic fibroblast growth factor, transforming growth factor-beta and beta-thromboglobulin.
Claim 6
Dependent Claim 29 words
A human in vitro cell line of claim 1, certain of said cells being morphologically and cytochemically similar to at least colony forming unit megakaryoblasts of the megakaryocytic series.
Claim 7
Dependent Claim 26 words
A human in vitro cell line of claim 1, wherein said cells undergo further differentiation to a more mature form when exposed to a phorbol ester.
Claim 8
Dependent Claim 41 words
A human in vitro cell line of claim 1, wherein said cells have the ability to express at least one protein selected from the group consisting of gpIIbIIIa protein complex, GMP-140, platelet Ca ++ ATPase, CD13, CD33, CD38, OKT9 and HLA-DR.
Claim 9
Dependent Claim 37 words
A human in vitro cell line of claim 1, certain of said cells having the ability to express at least one mRNA encoding a protein selected from the group consisting of PDGF-A chain, bFGF, TGF-beta and TGF-alpha.
Claim 10
Dependent Claim 20 words
A human in vitro cell line as deposited with the American Type Culture Collection (ATCC) under accession no. CRL 10107.
Claim 11
Dependent Claim 20 words
A human in vitro cell line as deposited with the American Type Culture Collection (ATCC) under accession no. CRL 10107.

References Cited (Backward Citations)

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USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1996-03-29 1996-09-28 1997-03-30 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 2000-03-29 2000-09-28 2001-03-30 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2004-03-29 2004-09-28 2005-03-30 $7700.00 Window Closed / Lapsed

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