大鼠脂肪干细胞冷冻复苏后的生物学特性及成骨细胞分化潜能的研究

Biological characteristics and osteogenic differentiation potential of adipose tissue derived stem cell

摘要:

目的 研究冻存复苏后大鼠脂肪干细胞（ADSCs）的生物学特性及其体外诱导成骨细胞分化的潜能。方法 冷冻保存6 个月的大鼠ADSCs 复苏后传代培养，显微镜下观察细胞形态；CCK8 检测细胞增殖，流式细胞仪检测CD44，CD105。第3 代细胞诱导成骨培养2 ～ 3 周后碱性磷酸酶（ALP）和茜素红染色，观察成骨情况。结果 复苏后ADSCs 形态类似成纤维细胞，增殖迅速。成骨诱导培养后表现出成骨细胞特性，ALP 染色活性增加，茜素红染色出现矿化结节。结论 冷冻保存复苏后的ADSCs 细胞生物性能稳定，定向诱导后可分化为 成骨细胞，可作为骨组织工程的种子细胞。

Abstract:

Objective To investigate biological characteristics of rat adipose derived stem cells (ADSCs) after cryopreservation and its osteogenic differentiation potential. Methods Rat ADSCs were revived and cultured.The morphological changes were observed under microscope. Cell proliferation were determined by CCK8. Flow cytometry was performed to measure CD44 and CD105 expression. In the 3rd generation of ADSCs, cells were induced for 2-3 weeks for osteogenesis. Osteogenic differentiation was determined by measuring expression of alkaline phosphates and Alizarin Red. Results ADSCs were successfully revived following cryopreservation,morphologically similar as fibroblast with fast proliferating capacity. Osteogenesis induction led to increased expression of ALP. Alizarin Red staining indicated mineralized nodules. Conclusion ADSCs functions well after recovery from cryopreservation and maintain differential potential to osteocyte, which suggests that ADSCs may be acceptable material for bone engineering.