Document Detail


Human pulp-derived cells immortalized with Simian Virus 40 T-antigen.
MedLine Citation:
PMID:  16630306     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Kerstin M Galler; Helmut Schweikl; Birger Thonemann; Rena N D'Souza; Gottfried Schmalz
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  European journal of oral sciences     Volume:  114     ISSN:  0909-8836     ISO Abbreviation:  Eur. J. Oral Sci.     Publication Date:  2006 Apr 
Date Detail:
Created Date:  2006-04-24     Completed Date:  2006-06-06     Revised Date:  2010-02-05    
Medline Journal Info:
Nlm Unique ID:  9504563     Medline TA:  Eur J Oral Sci     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  138-46     Citation Subset:  D; IM    
Affiliation:
Department of Operative Dentistry and Periodontology, University of Regensburg, Germany. kerstin.galler@uth.tmc.edu
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MeSH Terms
Descriptor/Qualifier:
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:
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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