小型猪ASCs生物学性状研究及其骨组织工程应用探讨

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。

Study on the Characteristics of Miniature Pig Adipose Derived Stem/Stromal Cells and its Application in Bone Tissue Engineering

Objective:To observe the morphological and biological characteristics and mutilineage potential of adipose derived stem/stromal cells (ASCs) of miniature pig. Otherwise, we discussed its application and experimental study in bone tissue engineering.Methods:ASCs were isolated from the subcutaneous adipose tissue of miniature pig by collagenase digestion. The characteristics and morphology of ASCs were observed by microscopic observation, flow cytometry, and lipogenic and osteogenic differentiation. The differentiations were identified by Oil Red O and Alizarin red staining respectively.Results:ASCs appeared to be spindle-shaped, a general bionomics of ASCs, and expressed positive of CD29 and CD90, while negative of CD14, CD31 and CD34. Moreover, ASCs were able to differentiate into adipocytes and osteoblasts.Conclusion:It is simple and feasible to obtain enough adipose tissue from the back of miniature pig, and fromwhich ASCs can be effectively isolated. Besides, ASCs have good proliferation and multipotential ability.