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背景:实验证实鹿茸多肽可以促进体外培养软骨细胞的增殖和细胞外基质糖胺多糖、Ⅱ型胶原、Aggrecan蛋白的表达。目的:通过对体外培养的兔骨髓间充质干细胞在特定培养液作用下向软骨细胞表型分化的研究,探讨鹿茸多肽对其软骨分化的影响。方法:将第3代兔骨髓间充质干细胞随机分为空白对照组、诱导组、鹿茸多肽组,分别采用普通培养液、诱导培养液、含10mg/L鹿茸多肽的诱导培养液于离心管内进行培养;并取兔的关节软骨细胞作为关节软骨组。分别于1,2,3周后取材,通过组织学、生物化学和RT-PCR技术,对离心管内构建的软骨组织进行形态学和细胞功能状态的观察。结果与结论:空白对照组培养2周后,细胞团块逐渐崩解,无法进行苏木精-伊红染色。诱导组、鹿茸多肽组细胞团块除有轻度收缩外,呈白色半透明状;苏木精-伊红染色发现部分细胞为圆形或卵圆形,表层细胞密度大;诱导组、鹿茸多肽组糖胺多糖含量及Ⅱ型胶原mRNA表达随培养时间延长而增多,各时间点诱导组、鹿茸多肽组含量均高于空白对照组(P<0.05);各时间点鹿茸多肽组糖胺多糖含量及Ⅱ型胶原mRNA表达均高于诱导组,但低于关节软骨组(P<0.05)。提示骨髓间充质干细胞在特定培养条件下能向软骨细胞表型分化,且鹿茸多肽对其定向软骨分化有明显促进作用。虽然在体外可以构建出软骨组织,但其与关节软骨质量相比仍有很大差距。
BACKGROUND: Experiments demonstrate that pilose antler polypeptides can promote the proliferation of chondrocytes cultured in vitro and the expression of extracellular matrix glycosaminoglycans, type Ⅱ collagen and Aggrecan protein. OBJECTIVE: To study the phenotypic differentiation of rabbit bone marrow mesenchymal stem cells cultured in vitro in specific culture medium to explore the effect of pilose antler polypeptide on the differentiation of chondrocytes. Methods: The third generation of rabbit bone marrow mesenchymal stem cells were randomly divided into blank control group, induction group and pilose antler polypeptide group, respectively, using ordinary culture medium, induced medium, containing 10mg / L pilose antler polypeptide culture medium in centrifuge tube Culture; and take rabbit articular chondrocytes as articular cartilage group. After 1, 2 and 3 weeks respectively, the morphological and cellular functions of cartilage tissue were observed by histological, biochemical and RT-PCR techniques. RESULTS AND CONCLUSION: After 2 weeks of culture in control group, cell mass gradually disintegrated and hematoxylin-eosin staining could not be performed. In the induced group and the antler polypeptide group, the cell mass was translucent except for mild contraction; some cells were round or oval in the hematoxylin-eosin staining, The content of glycosaminoglycan and the expression of type Ⅱ collagen mRNA increased with the prolongation of culture time, and the contents of induction group and antler polypeptide group were higher than those of blank control group at each time point (P <0.05). The content of glycosaminoglycan And type Ⅱ collagen mRNA expression were higher than the induction group, but lower than the articular cartilage group (P <0.05). These results suggest that MSCs can differentiate into chondrocytes under specific culture conditions and that pilose antler polypeptides can significantly promote the differentiation of their targeted cartilage. Although cartilage tissue can be constructed in vitro, there is still a large gap between it and the quality of articular cartilage.