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目的从兔肋软骨膜中分离间充质干细胞(Mesenchymal stem cells,MSCs),并进行鉴定。方法取3～4周龄新西兰大白兔,采用机械消化法和酶消化法结合的两步消化法获取肋软骨膜细胞,体外培养观察其生长形态;免疫组化法鉴定其表型特征;并检测其诱导成骨、成脂的分化潜能。结果从肋软骨膜中分离的原代细胞聚集成团,以圆形和多角形为主,从第2代开始细胞形态均一,以梭形为主,呈漩涡状排列,并开始表达CD29和CD90,不表达CD34和Ⅱ型胶原,现已传至15代,生长良好;第3代细胞经诱导可分化为成骨细胞或脂肪细胞。结论从兔肋软骨膜中可获得干细胞,其具有与MSCs相似的特性。
Objective To isolate and identify mesenchymal stem cells (MSCs) from rabbit costal cartilage. Methods New Zealand white rabbits (3 ~ 4 weeks old) were used to obtain the perichondrial pericytes cells by two-step digestion with mechanical digestion and enzymatic digestion. The growth of the cells was observed in vitro. The phenotypic characteristics were identified by immunohistochemistry. It induces osteogenic, adipogenic differentiation potential. Results The primary cells separated from the perichondrial periosteum aggregated into clusters with round and polygonal shapes. From the second generation, the cells were homogeneous and dominated by fusiform pattern with swirling arrangement and began to express CD29 and CD90 , Does not express CD34 and type Ⅱ collagen, has been passed to 15 generations, well-grown; 3rd generation of cells can be induced to differentiate into osteoblasts or adipocytes. Conclusion Stem cells can be obtained from rabbit costal cartilage and have similar characteristics to MSCs.