冻存SD大鼠脂肪组织中脂肪干细胞的分离培养和鉴定  

Isolation,culture and identification of adipose-derived stem cells from SD rat adipose tissues subjected to long-term cryopreservation

作　　者：刘琴[1] 王丽平[1] 陈芳[1] 张宜[1] LIU Qin WANG Liping CHEN Fang ZHANG Yi(Department of Medical Experiments, Wuhan General Hospital of Chinese PLA, Wuhan 430070, Chin)

机构地区：[1]中国人民解放军武汉总医院医学实验科,湖北武汉430070

出　　处：《细胞与分子免疫学杂志》2017年第2期202-205,共4页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的研究从冻存SD大鼠脂肪组织中分离培养脂肪干细胞(ADSC)的可行性。方法取健康SD大鼠股沟处脂肪组织,用100 m L/L二甲基亚砜联合900 m L/L胎牛血清(FBS)作为冻存剂,冻存于液氮中。3个月后复苏冻存的脂肪组织,进行ADSC的分离培养,观察细胞的生长状况及形态特征。取第3代细胞,CCK-8法绘制其生长曲线,免疫荧光技术检测CD29、CD45、CD90的表达;分别用成脂、成骨诱导培养液进行诱导分化,油红O、茜素红染色鉴定细胞分化情况。结果从冻存SD大鼠脂肪组织中分离得到的ADSC形态为长梭形纤维样,具有较好的增殖能力,生长曲线为典型的"S"型;免疫荧光技术证实细胞强表达CD29、CD90,CD45阴性;分离的细胞经成脂诱导后油红O染色为阳性,经成骨诱导后茜素红染色为阳性。结论从冻存SD大鼠脂肪组织中能够分离出ADSC。Objective To study the feasibility of isolation and culture of adipose-derived stem cells（ ADSCs） from SD rat adipose tissues subjected to long-term cryopreservation. Methods We took inguinal fat pads from healthy SD rats. Adipose tissues were stored with 100 m L / L dimethyl sulfoxide（ DMSO） combined with 900 m L / L fetal bovine serum（ FBS） in liquid nitrogen. Three months later,the adipose tissues were resuscitated for the isolation and culture of ADSCs. The growth status and morphology were observed. The growth curve and cell surface markers CD29,CD45,CD90 of the 3rd passage cells were analyzed respectively by CCK-8 assay and immunocytochemistry. The 3rd passage cells were induced towards adipogenic lineages and osteogenic lineages by different inducers,and the resulting cells were examined separately by oil red O staining and alizarin red staining. Results The ADSCs obtained from SD rat adipose tissues subjected to long-term cryopreservation showed a spindle-shape appearance and had a good proliferation ability. The cell growth curve was typical ＂S ＂ curve.Immunocytochemistry showed that the 3rd passage cells were positive for CD29 and CD90,while negative for CD45. The cells were positive for oil red O staining after adipogenic induction,and also positive for alizarin red staining after osteogenic induction. Conclusion The ADSCs can be isolated from SD rat adipose tissues subjected to long-term cryopreservation.

关 键 词：SD大鼠 脂肪组织 冻存 脂肪干细胞(ADSC) 

分 类 号：R392-33[医药卫生—免疫学] Q254[医药卫生—基础医学]