Beagle犬脂肪基质干细胞的培养及分化研究

Primary culture and differentiation of beagle dog adipose-derived stromal cells

目的:体外培养Beagle犬脂肪基质干细胞(dog adipose-derived stromal cells,dADSCs),定向诱导分化为脂肪、成骨细胞,为口腔组织工程提供种子细胞。方法:取Beagle犬皮下脂肪组织,剪碎,经I型胶原酶消化培养犬脂肪基质干细胞,取第3代细胞鉴定其来源及干细胞标记并向脂肪、成骨细胞诱导分化。结果:该细胞波形丝蛋白表达阳性,角蛋白表达阴性,CD44表达阳性;成脂诱导后,经油红O染色可见细胞内脂滴的积累;成骨诱导后,ALP、钙结节、骨钙素、I型胶原均阳性表达。结论:Beagle犬脂肪组织中存在着具有能分化为脂肪、成骨细胞的脂肪基质干细胞,此脂肪基质干细胞可作为口腔组织工程种子细胞来源。

Objective： To induce the differentiation of beagle dog adipose-derived stromal cells to adipocyte, osteoblast for oral tissue engineering. Methods： Digested by collagenase Ⅰ, passage 3 adipose-derived stromal cells from Beagle dog abdomen subcutaneous adipose tissues were cultured and identified and induced into adipocytes, osteoblast. Results： The cells were Vimentin （ ＋ ）, CD44（ ＋ ） and keratin （ - ）. Stained by Oil Red O, differentiated adipocytes had accumulation of lipid droplets. After osteogenic induction, the cells were ALP （ ＋ ）, osteocalcin （ ＋ ）, collagen 1 （ ＋ ） and positively stained by alizarin Bordeaux. Conclusion ： In matured adipocyte tissue there are adipose-derived stromal cells with potential ability of orientation differentiation into adipocytes, osteoblasts by suitable induction. The results suggest that the dASCs may be the ideal cells for oral tissue engineering in future.