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目的:观察小型猪脂肪来源干细胞(adipose derived stem/stromal cells,ASCs)的形态学特点,研究其多向分化潜能和生物学特性,探讨其在骨组织工程方面的应用。方法:切取雄性小型猪背部皮下脂肪,体外分离培养并鉴定ASCs,观察其生长、增殖特性和组织学形态;并诱导其向脂肪细胞和骨细胞多向分化,分别采用油红O染色和茜素红染色鉴定,细胞表面分子通过流式细胞术鉴定。结果:贴壁的ASCs为长梭形,生长增殖快,性状稳定,流式细胞术检测显示CD29、CD90阳性表达,CD14、CD31、CD34阴性表达。通过相应的诱导培养液诱导,ASCs可向脂肪细胞和骨细胞分化并表现出相应的生物学特征。结论:小型猪背部皮下脂肪取材方便,可获得大量脂肪用以分离ASCs,ASCs增殖分化能力较强,在条件培养基诱导下可成骨分化,可作为种子细胞应用于骨组织工程。
OBJECTIVE: To observe the morphological characteristics of adipose derived stem / stromal cells (ASCs) and to investigate their multi-directional differentiation potential and biological characteristics and to explore their applications in bone tissue engineering. Methods: Subcutaneous adipose tissue was isolated from the back of male piglets. ASCs were isolated and cultured in vitro. The growth, proliferation and histomorphology of ASCs were observed. The adipocytes and osteoblasts were induced to differentiate into adipocytes and osteoblasts. Oil red O staining and alizarin Identification by red staining, identification of cell surface molecules by flow cytometry. Results: The adherent ASCs were long fusiform, with fast growth and stable proliferation. The results of flow cytometry showed that CD29 and CD90 were positive and CD14, CD31 and CD34 were negative. Induced by the corresponding culture medium, ASCs can differentiate into adipocytes and osteoblasts and exhibit corresponding biological characteristics. CONCLUSION: The subcutaneous fat in the back of miniature pig is easy to be obtained, and a large amount of fat can be obtained for the isolation of ASCs. The proliferation and differentiation of ASCs is strong. Under the condition of medium induction, osteogenic differentiation of ASCs can be used as seed cells in bone tissue engineering.