| Immortalization of human articular chondrocytes for generation of stable, differentiated cell lines. | |
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MedLine Citation:
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PMID: 15280585 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Immortalized chondrocytes of human origin have been developed to serve as reproducible models for studying chondrocyte function. In this chapter, methods for immortalization of primary human chondrocytes with SV40-TAg, HPV-16 E6/E7, and telomerase by retrovirally mediated transduction and selection for neomycin resistance are described. However, stable integration of an immortalizing gene stabilizes proliferative capacity, but not the differentiated chondrocyte phenotype. Thus, strategies for selection of chondrocyte cell lines, involving the maintenance of high cell density and moderation of cell proliferation, are also described. The methods for immortalization and selection are applicable to the development of chondrocyte cell lines using any immortalizing agent. Although immortalized chondrocytes should not be considered as substitutes for primary chondrocytes, they may be useful tools for evaluating and further validating mechanisms relevant to cartilage biology. |
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Authors:
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Mary B Goldring |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Methods in molecular medicine Volume: 100 ISSN: 1543-1894 ISO Abbreviation: Methods Mol. Med. Publication Date: 2004 |
Date Detail:
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Created Date: 2004-07-28 Completed Date: 2005-01-31 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 101123138 Medline TA: Methods Mol Med Country: United States |
Other Details:
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Languages: eng Pagination: 23-36 Citation Subset: IM |
Affiliation:
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Division of Rheumatology, Beth Israel Deaconess Medical Center, New England Baptist Bone and Joint Institute, Harvard Institutes of Medicine, Boston, MA, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Cartilage, Articular
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cytology* Cell Culture Techniques Cell Differentiation Cell Line, Transformed* Cell Transformation, Viral / genetics Chondrocytes / cytology*, virology Extracellular Matrix Proteins / genetics Gene Expression Humans Oncogene Proteins, Viral / genetics RNA, Messenger / analysis Repressor Proteins / genetics Retroviridae / genetics Simian virus 40 / genetics Telomerase / genetics |
| Grant Support | |
ID/Acronym/Agency:
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AG 22021/AG/NIA NIH HHS; AR 45378/AR/NIAMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/E6 protein, Human papillomavirus type 16; 0/Extracellular Matrix Proteins; 0/Oncogene Proteins, Viral; 0/RNA, Messenger; 0/Repressor Proteins; EC 2.7.7.49/Telomerase |
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