兔脂肪干细胞体外成骨诱导培养及成骨活性鉴定的实验研究

目的体外分离培养兔脂肪干细胞（adipose-derived stem cells,ADSCs）,在成骨诱导条件下鉴定其成骨活性。方法取4月龄新西兰大白兔腹股沟区脂肪,Ⅰ型胶原酶消化,分离、培养及传代原细胞,将第3代ADSCs消化后,加入成骨培养液中诱导分化,分别行碱性磷酸酶染色、茜素红染色、VonKoss（a钙结节）染色,以鉴定分化结果。结果体外分离培养的细胞增殖活跃,成骨诱导后碱性磷酸酶染色、茜素红染色及Von Kossa染色均呈阳性表达。结论脂肪干细胞具有成骨分化能力,是一种理想的骨组织工程种子细胞。

An experimental study on osteoinductive culture in vitro and osteogenic activity identification of rabbit adipose-derived stem cells

Objective To Separate and cultivate rabbit adipose-derived stem cells （ADSCs） and identify its osteogenic activity under osteoinductive conditions. Methods Inguinal fat extracted from New Zealand white rabbits aged 4 months was digested by collagenase Ⅰ, primary cells were separated, cultured and passaged. Thereafter, the third generation ADSCs were digested, and then induced and differentiated by adding osteogenic culture solution. Alkaline phosphatase （ALP） staining, alizarin red staining and Von Kossa （calcium nodules） staining were carried out to identify the differentiation. Results ADSCs proliferation was active, the results of stainings of ALP, alizarin red and Von Kossa showed positive expression after osteoinduction. Conclusion ADSCs can be an ideal seed cell for bone tissue engineering due to its osteogenic differentiation potential.