目的体外分离培养大鼠脂肪源性干细胞并进行鉴定,为脂肪源性干细胞移植治疗勃起功能障碍大鼠模型提供良好的种子细胞。方法Ⅰ型胶原酶法原代培养大鼠脂肪源性干细胞。取第4代细胞,流式细胞术检测间充质干细胞表面分子CD31、CD34、CD45、CD73、CD90和CD105,并采用成脂、成骨、成平滑肌和成内皮诱导培养基多向诱导分化后通过油红O染色、茜素红染色、免疫荧光检测α-平滑肌肌动蛋白(α-SMA)和血管性血友病因子(vWF)进行鉴定。结果所培养的细胞CD73、CD90和CD105表达阳性,而CD31、CD34和CD45表达阴性。多向诱导分化后,油红O染色、茜素红染色、免疫荧光检测α-SMA和vWF均为阳性,而对照组为阴性。结论成功从大鼠脂肪组织中分离培养脂肪源性干细胞,可用于勃起功能障碍的脂肪源性干细胞移植治疗。%Objective To culture and identify rat adipose-derived stem cells in vitro, and provide the available seed cells for erectile dysfunction therapy. Methods Adipose-derived stem cells were isolated and cultured from the inguinal region of SD rat. The expressions of CD31, CD34, CD45, CD73, CD90 and CD105 in the isolated stem cells were arrayed by flow cytometry. The fourth passage cells were induced and identified by their direct differentiation capacities to the adipogenic, osteogenic, myogenic and endothelialcells. Results The growth curve of the fourth passage cells presented typical reversed“S”. The cultured cells expressed CD73, CD90 and CD105, lacked CD31, CD34 and CD45. The differentiated adipogenic, Osteogenic, Myogenic and endothelial differentiation cells were identified by Oil Red staining, Alizarine Red staining, α-SMA expression and vWF expression, respectively. Conclusion Rat adipose-derived stem cells have been successfully isolated and cultured, and could be used as autogenous adipose-derived stem cells for erectile dysfunction therapy.
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