兔脂肪间充质干细胞的生物学特性及其成骨诱导分化的研究  

Biological characteristics of rabbit adipose-derived stem cells and their induction to osteogenesis

作　　者：谢永辉[1] 陈庄洪[2] 蔡贤华[2] 王清富[1] 王华松[2] 刁波[3] 

机构地区：[1]南方医科大学,广州510515 [2]广州军区武汉总医院骨科,广州510515 [3]广州军区武汉总医院医学科,广州510515

出　　处：《中华实验外科杂志》2013年第2期358-360,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨脂肪问充质干细胞的分离培养方法，观察其生物学特性及成骨分化潜能。方法将成年日本大耳白兔的腹股沟白色脂肪组织分离取出，通过1％I型胶原酶消化逐步提取脂肪问充质干细胞，待细胞融合约80％时传代，定期用倒置显微镜观察细胞间充质干细胞的超微结构、表面分子标志表达，收集第3、5、10、15代细胞，描绘其生长曲线，加入成骨诱导培养液进行诱导分化并观察。结果脂肪问充质干细胞各代的生长曲线无明显差异，均成“s”形。Gomori萘酚磷酸酯法染色显示其胞质内富含碱性磷酸酶颗粒、VonKossa染色显示聚集的细胞团能形成矿化结节。脂肪间充质干细胞表面标志检测结果为：CD29和CD44阳性，CD34和CD45阴性。结论脂肪间充质干细胞来源广泛、含量丰富、生物学性能稳定，具有成骨分化潜能。Objective To investigate the isolation and culture of adipose-derived stromal cells （ADSCs） and observe the biological characteristics and osteogenic differentiation potential of ADSCs. Methods The white adipose tissue was obtained from the inguinal fat pads of adult Japanese white rab- bits, and digested with type I collagenase. Primary ADSCs were passaged and cultured when the cells were about 80% confluent. The ultrastructure and surface marker expression of ADSCs were observed under the inverted microscopy. The growth curve was made from the 3rd, 5th, 10th, and 15th generations of ceils. The possibility of inducing ADSCs to osteogenesis in specific differentiation medium was observed. Results The growth curves of generations of cells had no significant diffenence, which displayed reversed S-shaped. Gomori and Von Kossa staining demonstrated postitive expression of ALP and formation of miner- alized nods in extracellular matrix. The cultured ADSCs were positive for CD29 and CD44, and negative for CD34 and CD45. Conclusion The abundant ADSCs can be induced to differentiate into osteogenesis, and have stable biological characteristics. ADSCs may be served as seeding cells in the bone tissue engineer- ing.

关 键 词：脂肪干细胞 培养 成骨 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]