摘要
目的探索大鼠脂肪组织来源的干细胞的分离、培养的方法,以及在特定条件下向成骨细胞分化,探讨其作为骨组织工程种子细胞的可行性。方法取4周龄SD大鼠附睾处脂肪,成骨诱导培养基培养,组织化学、免疫细胞化学染色以及组织学定量检测其分化情况。结果从成体大鼠脂肪组织中培养出脂肪组织来源的干细胞,能稳定增殖传代,CD44、波形蛋白免疫组化染色阳性,证明其中胚层来源。在地塞米松、维生素C、β-甘油磷酸钠的诱导下,脂肪组织来源干细胞的碱性磷酸酶活性增高,出现钙结节,四环素荧光染色阳性证实有新骨形成。碱性磷酸酶定量和钙定量随诱导时间延长表达增加。结论从脂肪组织中可获得具有多向分化潜能的干细胞,并能在体外稳定增殖传代,经诱导后可分化为成骨细胞,有可能成为骨组织工程较理想的种子细胞之一。
Objective To investigate the possibility of differentiation of adipose tissue-derived stromal cells into osteoblasts in vitro. Methods Adipose tissue was obtained from epididymis of SD rat with collagenase digestion. And the stromal cells from adipose tissue （ATSCs） were cultured and differentiated into osteoblasts induced by dexamethasone, vitamin C and phosphoglycerol. The differentiated cells were identified by immunochemical staining and cytochemistrical examination. Results ATSCs were successfully isolated from rat adipose tissue, and they were positively stained for CD44 and Vimentin. They appeared fibroblast-like cells and could be stably proliferated in vitro. After treatment with dexamethasone, ascorbic acid and beta-sodium glycerolphosphate, the differentiation of ATSCs towards osteoblasts was verified by the positive staining of ALP, and the levels of ALP and calcium produced by the differentiated cells were gradually increased. Conclusions ATSCs obtained from rat adipose tissue can differentiate to osteoblasts in vitro, and they may be served as a novel autogenous cell source for bone tissue engineering.
出处
《中华小儿外科杂志》
CSCD
北大核心
2006年第7期379-382,共4页
Chinese Journal of Pediatric Surgery
基金
江苏省卫生厅135工程课题资助(38RC2002038)
