兔脂肪干细胞的分离培养鉴定及成骨诱导分化研究

Osteogenic differentiation of adipose-derived stem cells from rabbits in vitro

[目的]探讨在成骨诱导条件下兔脂肪干细胞的体外诱导分化情况。[方法]取3个月龄日本大耳白兔颈背部皮下脂肪,用Ⅰ型胶原酶消化获得细胞。Stro-1免疫细胞化学染色鉴定细胞性质;加入成骨诱导液后依次进行形态学、Ⅰ型胶原、碱性磷酸酶及钙盐沉积相关检测。[结果](1)原代所获细胞Stro-1表达阳性。(2)在诱导条件下,Ⅰ型胶原、碱性磷酸酶及钙盐沉积均呈阳性表达。[结论]脂肪干细胞来源广、易获取、对机体创伤小,与骨髓间充质干细胞类似,经体外诱导后可实现成骨分化,为骨组织工程提供了一种新的种子细胞。

[ Objective] To observe the differentiation of adipose derived stem cells （ADSCs） into osteogenic cells in rabbit in vitro. [ Method] ADSCs were prepared by collagenase Ⅰ digestion of subcutaneous fat from the nape of Japanese white rabbit ＇after being excised and finely minced. Cells were identified by stro-1 immunocy to chemistry, and collagen Ⅰ immunocytochemistty, alkaline phosphatase assay and Von Kossa stain after osteogenic differentiation was perforlned. [ Result] （ 1 ） ADSCs were stro-1 positive. （2） The expression of collagen Ⅰ , alkaline phosphatase and caMum deposit of ADSCs were all positive under the influence of osteogenic differentiation medium. [ Conclusion] ADSCs have similar characteristics to bone marrow stromal cells, but are much more easier to have high number upon harvest and bring less pain will occur during taking ADSCs than taking BMDCs.