Document Detail


Phenotypic characterization of the 3/A/1D-1M osteogenic cell line derived from in vivo transplantation of 3/A/1D-1 chondroprogenitor murine teratocarcinoma cells.
MedLine Citation:
PMID:  8855376     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Bone cells involved in the replacement of cartilage by bone in the endochondral ossification process are known to enter via the medullar pathway. A hypothesis for the development of osteoblasts from chondroblasts was investigated by analyzing the phenotypic characteristics of the 3/A/1D-1M cell line derived from endochondral bone ossicle which was formed after in vivo transplantation of 3/A/1D-1 chondroprogenitor mouse teratocarcinoma cells. The 3/A/1D-1M cell cultures exhibited a triphasic evolution: after reaching confluence (day 3), cultures developed well-delimited cell clusters (days 6-8), which ultimately were organized into multilayered nodules (days 12-15). Electron-microscopic examination of such nodules at day 18 showed the presence of needle-shaped crystals associated with collagen fibrils in the extracellular space. The kinetics of collagen expression, investigated by an immunofluorescence staining procedure showed that, while confluent cultures mainly expressed type III collagen (70% of cells) with some type I (30-40% of cells) and V (30-40% of cells), the type I collagen became the major isoform beginning with day 6. From day 6 onwards, NP40-extracted alkaline phosphatase (AP) activity appeared concomitantly to cell cluster formation, and reached 160 nmol/min/mg of protein at the stage of nodule maturation (day 15). The strong inhibition of enzymatic activity by levamisole and L-homoarginine (IC50 = 0.9 microM and 5 mM, respectively) and its rapid heat inactivation at 56 degrees C (IT50 = 90 s), revealed the bone specificity of AP expressed by 3/A/1D-1M cells. In confluent cultures, brief exposure to parathyroid hormone (10 nM), known to be a bone-resorbing agent, showed a 60% increase in the intracellular cAMP level. In addition, while producing mRNA for the bone-specific protein osteocalcin, 3/A/1D-1M cells also produced type II procollagen mRNA, known to be the major cartilage-related characteristic. This in vitro study demonstrates that the 3/A/1D-1M clonal cell line, originating from 3/A/1D-1 chondroprogenitor cells after in vivo passage, was able to develop differentiated osteoblastic properties as well as the residual expression of the major chondrocytic RNA messenger.
Authors:
A Cohen-Tanugi; M Bolle; M L Boy-Lefèvre; F Anagnostou; N Forest
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Differentiation; research in biological diversity     Volume:  60     ISSN:  0301-4681     ISO Abbreviation:  Differentiation     Publication Date:  1996 Sep 
Date Detail:
Created Date:  1996-12-09     Completed Date:  1996-12-09     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  0401650     Medline TA:  Differentiation     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  327-37     Citation Subset:  IM    
Affiliation:
Laboratoire de Biologie-Odontologie, Université Paris 7, France.
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MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / biosynthesis
Animals
Cartilage / cytology,  metabolism
Cell Differentiation
Cell Line
Clone Cells
Collagen / biosynthesis
Cyclic AMP / biosynthesis
Extracellular Matrix / metabolism
Gene Expression Regulation, Neoplastic
Mice
Neoplasm Transplantation
Neoplasms, Experimental / drug therapy,  metabolism,  pathology
Osteoblasts / cytology*
Osteocalcin / genetics
Parathyroid Hormone / pharmacology
Phenotype
Procollagen / genetics
RNA, Messenger / biosynthesis
Rats
Stem Cells / cytology*
Teratocarcinoma / drug therapy,  pathology*
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Parathyroid Hormone; 0/Procollagen; 0/RNA, Messenger; 104982-03-8/Osteocalcin; 60-92-4/Cyclic AMP; 9007-34-5/Collagen; EC 3.1.3.1/Alkaline Phosphatase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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