Culture of rabbit bone marrow mesenchymal stem cells and collagen scaffolds under the mimic microgravity environment in vitro
Chen, C., et al. (2010). "Culture of rabbit bone marrow mesenchymal stem cells and collagen scaffolds under the mimic microgravity environment in vitro." Journal of Clinical Rehabilitative Tissue Engineering Research 14 34: 6308-6312
BACKGROUND:The method of differentiating from bone marrow mesenchymal stem cells(BMSCs) into chondrocytes included in vitro high-density micelle culture,in vitro monolayer cell culture,in vitro three dimensional stent induction,in vitro co-culture induction with chondrocytes and gene transfection.OBJECTIVE:To study adhesion,extension and proliferation of rabbit BMSCs in type Ⅰ and Ⅱ collagen scaffolds under the mimic microgravity environment in vitro.METHODS:BMSCs of rabbits were primarily cultured and subcultured in vitro,and then divided into two groups according to the difference of induction factors:experimental group receiving transforming growth factor(TGF)-β1 and insulin-like growth factor(IGF)-Ⅰ ;blank control group.After three weeks,the two groups were detected by methyl thiazolyl tetrazolium(MTT) assay,measurement of glycosaminoglycan(GAG) and immunohistochemistry.Chondrocytes in the experimental group were incubated in type Ⅰ and Ⅱ collagen scaffolds,and then divided into four groups:group 1:chondrocytes and type Ⅱ collagen scaffolds co-cultured statically;group 2:chondrocytes and type Ⅱ collagen scaffolds co-cultured under the mimic microgravity environment;group 3:chondrocytes and type Ⅰ collagen scaffolds co-cultured statically;group 4:chondrocytes and type Ⅰ collagen scaffolds co-cultured under the mimic microgravity environment.One week later,hematoxylin-eosin staining and toluidine blue staining were performed.RESULTS AND CONCLUSION:The results of the MTT assay(absorbance value) and the GAG content in experimental group were higher than in blank control group.Immunohistochemical detection of collagen Ⅱ was positive in experimental group.Results from hematoxylin-eosin staining and toluidine blue staining have demonstrated that composited cell number in type Ⅱ collagen scaffolds was evidently more than that of type Ⅰ collagen.Composited cell number under the mimic microgravity environment was evidently more than that of static culture in the same scaffold.Above-described results have confirmed that the mimic microgravity environment is conducive to adhesion and proliferation of high-density cells,and contributes to signal transmission among cells to provide suitable microenvironment for maintaining cell growth and metabolism.Collagen type Ⅱ scaffold can be used as a more satisfying scaffold material for chondrocytes than collagen type Ⅰ
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