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目的:研究大鼠脂肪基质干细胞(adipose-derived stem cells,ADSCs)分离培养的方法,探讨大鼠脂肪基质干细胞在体外向成骨细胞分化的能力。方法:取成年Sprague-Dawley大鼠腹股沟处脂肪组织,胶原酶消化分离,接种于自制基础培养基中分离传代。于基础培养基中传至第二代时改换诱导培养基,诱导向成骨细胞分化,培养2～4周,用碱性磷酸酶染色和Von Kossa染色鉴定成骨细胞分化能力。结果:大鼠脂肪中能够分离培养出生长旺盛的脂肪基质干细胞;经向成骨细胞诱导培养后,碱性磷酸酶染色和Von Kossa染色阳性,证实细胞能够分化为成骨细胞。结论:大鼠脂肪组织可分离培养出脂肪基质干细胞,生物学特性与骨髓基质干细胞(mes-enchymal stem cells,MSCs)相似,能够向成骨细胞分化,有望成为组织工程的种子细胞来源。
OBJECTIVE: To study the isolation and culture of rat adipose-derived stem cells (ADSCs) and investigate the ability of rat adipose-derived stem cells to differentiate into osteoblasts in vitro. Methods: The adipose tissue of adult Sprague-Dawley rats was digested by collagenase and seperated in homemade basal medium. When basal medium was transferred to the second generation, the induction medium was changed and osteoblast differentiation was induced and cultured for 2 to 4 weeks. Osteoblast differentiation ability was identified by alkaline phosphatase staining and Von Kossa staining. RESULTS: Rat adipose tissue-derived adipose-derived stem cells were isolated and cultured. After osteoblast induction, alkaline phosphatase staining and Von Kossa staining showed that the cells could differentiate into osteoblasts. CONCLUSION: Adipose-derived stem cells can be isolated and cultured in adipose tissue of rats. The biological characteristics are similar to that of mes-enchymal stem cells (MSCs). They can differentiate into osteoblasts and are expected to be the source of seed cells for tissue engineering.