同济大学学报（医学版）2012，Vol.33Issue(2)：1-6,6.DOI:10.3969/j.issn1008-0392.2012.02.001
犬成肌细胞体外向纤维软骨细胞表型分化的实验研究
Phenotype transformation from canine skeletal myoblast to fibrochondrocyte in vitro
摘要
Abstract
Objective To induce the phenotype transformation of canine skeletal myoblast into fibrochondrocyte in vitro. Methods The muscle from the hindlimb of adult Beagle canine was obtained, from which skeletal myoblasts were isolated and harvested by mechanical decomposition and two-step enzyme digestion. The cultured skeletal myoblasts of the fourth passage were digested and cultured in a medium containing human cartilage-derived morphogenic protein-2 ( hCDMP-2) in a 24-well plate at a density of 2. 0 × 104/cm2 in the experimental group; while in the control group the DMEM-LG complete culture medium without hCDMP-2 was used. After 21 days of incubation the cellular morphology, secretion function and expression of type Ⅱ collagen in cultured cells were examined by HE and toluidine blue staining, and immunohistochemistry, respectively. The expression of collagen Ⅰ , Ⅱ and Sox9 mRNA was detected by reverse transcription-polymerase chain reaction ( RT-PCR). Results The skeletal myoblasts changed from a spindle-like appearance into a polygonal shape after induction. The extracellular matrix was positive stained by H-E and toluidine blue staining in experimental group, and the cells were positively stained by immunohistochemistry for type Ⅱ collagen. RT-PCR revealed that the expression of collagen Ⅰ , Ⅱ and Sox9 mRNA were significantly displayed in the experimental group. No positive expression was observed in the control group. Conclusion The canine skeletal myoblasts can be induced for phenotype transformation to chondrocy-tes by hCDMP-2 in vitro.关键词
成肌细胞/软骨分化/种子细胞/人源性软骨源性形态发生蛋白-2/软骨组织工程Key words
skeletal myoblasts/ chondrogenic differentiation/ seeding cells/ human cartilage-derived morphogenic protein-2/ cartilaginous tissue engineering分类
医药卫生引用本文复制引用
顾羊林,朱文辉,王予彬,卢亮宇,程晏,李靖龙..犬成肌细胞体外向纤维软骨细胞表型分化的实验研究[J].同济大学学报（医学版）,2012,33(2):1-6,6.基金项目
上海市自然科学基金(09ZR1425500) （09ZR1425500）
