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Studies on osteogenic differentiation of rat bone marrow stromal cells cultured in type I collagen gel by RT‐PCR analysis
Author(s) -
Taira M.,
Toyosawa S.,
Ijyuin N.,
Takahashi J.,
Araki Y.
Publication year - 2003
Publication title -
journal of oral rehabilitation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 93
eISSN - 1365-2842
pISSN - 0305-182X
DOI - 10.1046/j.1365-2842.2003.01170.x
Subject(s) - osteonectin , stromal cell , osteopontin , osteocalcin , bone sialoprotein , type i collagen , bone marrow , alkaline phosphatase , chemistry , cell culture , microbiology and biotechnology , endocrinology , medicine , biology , immunology , biochemistry , enzyme , genetics
summary The purpose of this investigation was to examine by reverse‐transcriptase polymerase chain reaction analysis the osteogenic differentiation of twice‐passaged Sprague–Dawley rat bone marrow stromal cells in type I collagen gel cultured for 3 weeks. Two culture media were used here, namely Dulbecco's modified Eagle (DME) medium supplemented with vitamin C [Dex (–)] and those with vitamin C, dexamethasone and β ‐glycerophosphate [Dex (+)]. Culture with Dex (–) medium in collagen gel for 3 weeks brought about the well‐developed cell network and middle‐stage osteogenic phenotype expression characterized by mRNA for alkaline phosphatase, osteonectin and osteopontin while those for bone sialo protein and osteocalcin were not detected. On the contrary, culture with Dex (+) medium in collagen gel for 3 weeks lead to necrosis of the cells. These results indicate that culture in collagen gel with Dex (–) DME medium containing vitamin C was useful for three‐dimensional culture and middle‐stage osteogenic differentiation of twice‐passaged bone marrow stromal cells. This study might contribute to tissue engineering therapy to fix bone and periodontal defects in the future.