Gene expression profile of bovine bone marrow mesenchymal stem cell during spontaneous chondrogenic differentiation in pellet culture system.
Bovine bone marrow mesenchymal stem cells (MSCs) cultured in condensate culture, spontaneous and independent for any external biostimulants, undergo chondrogenic differentiation. In the present study, the bovine MSC chondrogenesis pathway was studied by analyzing stage-specific gene expression using quantitative "Real Time" reverse transcriptase polymerase chain reaction (qRT-PCR). Results showed that bovine MSCs underwent complete chondrogenesis; the initial stage was characterized by expression. of sox9 messenger ribonucleic acid (mRNA), followed by high transcription of chondrocyte specific genes, collagen type II and IX, biglycan and cartilage oligomeric matrix protein, and the final prehypertrophic and/or hypertrophic stage was distinguished by increased expression of collagen type X. From day 7 to day 14 of differentiation increased mRNA expression of the transforming growth factors beta1 and beta2, basic fibroblast growth factor (FGF 2), bone morphogenic protein 6 (BMP 6), insulin-like growth factors 1, parathyroid hormone related peptide and indian hedgehog (Ihh) were detected. These results suggest that these well know chondrogenic growth factors may play a role in bovine chondrogenesis in autocrine and/or paracrine manner. On day 21 of the culture, FGF 2, BMP 6 and Ihh were highly expressed, compared to cells cultured in monolayer manner, which suggests a possible function in maintaining the terminal stage of differentiation. This data extends our knowledge about the unusual species-specific bovine MSC chondrogenesis, allowing us to define the phenotype of the differentiated cells. Furthermore, this study contributes to our in understanding of known chondrogenic-growth factors in autocrine and/or paracrine manner playing a role in the spontaneous differentiation.
Laboratory of Veterinary Surgery, Department of Clinical Science, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan. Darko.Bosnakovski@UTSouthwestern.edu
Darko
D
Bosnakovski
Morimichi
M
Mizuno
Gonhyung
G
Kim
Satoshi
S
Takagi
Masahiro
M
Okumur
Toru
T
Fujinag
D000818:Animals; D002417:Cattle / Q000235:genetics*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D020219:Chondrogenesis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics
D012333:RNA, Messenger