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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 explore the isolation and culture methods of adipose tissue-derived stem cells in rats and to differentiate into osteoblasts under specific conditions and to explore the feasibility of using them as bone tissue engineered seed cells. Methods Adipose at the epididymis of 4-week-old SD rats were cultured, osteoinductive medium was cultured, histochemistry, immunocytochemistry and histology were used to detect the differentiation. Results Adipose tissue - derived stem cells were cultured from adult rat adipose tissue and stably proliferated and passaged. CD44 and vimentin immunohistochemical staining were positive, indicating the origin of mesoderm. Under the induction of dexamethasone, vitamin C and β-glycerophosphate, the alkaline phosphatase activity of adipose tissue-derived stem cells increased, calcium nodules appeared, and positive staining with tetracycline confirmed new bone formation. Alkaline phosphatase and calcium quantification with prolonged induction of expression increased. Conclusion Stem cells with multidirectional differentiation potential can be obtained from adipose tissue, and can proliferate and pass on in vitro steadily. After being induced to differentiate into osteoblasts, they may be one of the ideal seed cells for bone tissue engineering.