| Human pulp-derived cells immortalized with Simian Virus 40 T-antigen. | |
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MedLine Citation:
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PMID: 16630306 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Primary cells in culture have a limited capacity to divide and soon reach a non-proliferative state. This cellular senescence limits the investigation of cells derived from human pulp concerning cellular pathways, gene regulation, mechanisms of dentin formation, or responses to material exposure. To overcome this problem, primary human pulp-derived cells were established and transfected with a plasmid containing coding sequences of Simian Virus 40 (SV40) large T-antigen. This resulted in the establishment of several cell clones showing an extension of life span. Expression of T-antigen transcripts and protein was verified by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. Primary human pulp cells were cultured until senescence (i.e. up to passage 7) and transfected cells could be cultured to passage 18 after transfection, when a cellular crisis with massive cell death occurred. One clone escaped from crisis and has been maintained in culture for 55 wk. Experiments were performed to characterize transfected cells in comparison to primary cells. Cell morphology and proliferation were analyzed, and expression of cell-specific gene transcripts and proteins (including collagen types I and III, alkaline phosphatase, bone sialoprotein, osteocalcin, and dentin sialophosphoprotein and dentin matrix protein I) was detected by RT-PCR and immunohistochemistry. Transfection of human pulp-derived cells resulted in an immortalized cell line retaining many of the phenotypic characteristics observed in primary cells. |
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Authors:
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Kerstin M Galler; Helmut Schweikl; Birger Thonemann; Rena N D'Souza; Gottfried Schmalz |
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Publication Detail:
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Type: Comparative Study; Journal Article |
Journal Detail:
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Title: European journal of oral sciences Volume: 114 ISSN: 0909-8836 ISO Abbreviation: Eur. J. Oral Sci. Publication Date: 2006 Apr |
Date Detail:
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Created Date: 2006-04-24 Completed Date: 2006-06-06 Revised Date: 2010-02-05 |
Medline Journal Info:
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Nlm Unique ID: 9504563 Medline TA: Eur J Oral Sci Country: Denmark |
Other Details:
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Languages: eng Pagination: 138-46 Citation Subset: D; IM |
Affiliation:
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Department of Operative Dentistry and Periodontology, University of Regensburg, Germany. kerstin.galler@uth.tmc.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adolescent Adult Alkaline Phosphatase / genetics Antigens, Viral, Tumor / genetics* Cell Aging / genetics* Cell Death / genetics Cell Line Cell Proliferation Cell Shape / genetics Cells, Cultured Clone Cells / cytology Collagen Type I / genetics Collagen Type III / genetics Dental Pulp / cytology* Extracellular Matrix Proteins / genetics Humans Osteocalcin / genetics Phosphoproteins / genetics Sialoglycoproteins / genetics Simian virus 40 / genetics, immunology* Transcription, Genetic / genetics Transfection* |
| Chemical | |
Reg. No./Substance:
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0/Antigens, Viral, Tumor; 0/Collagen Type I; 0/Collagen Type III; 0/DMP1 protein, human; 0/Extracellular Matrix Proteins; 0/Phosphoproteins; 0/Sialoglycoproteins; 0/dentin sialophosphoprotein; 0/integrin-binding sialoprotein; 104982-03-8/Osteocalcin; EC 3.1.3.1/Alkaline Phosphatase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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