冻存SD大鼠脂肪组织中脂肪干细胞的分离培养和鉴定

发布时间：2018-10-11 15:30

【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。
[Abstract]:Objective to study the feasibility of isolation and culture of adipose stem cell (ADSC) from adipose tissue of cryopreserved SD rats. Methods Adipose tissue in the groin of healthy SD rats was frozen with 100ml / L dimethyl sulfoxide (DMSO) combined with 900ml / L fetal bovine serum (FBS) as cryopreservation agent and frozen in liquid nitrogen. After 3 months of resuscitation, ADSC was isolated and cultured. The growth and morphological characteristics of the cells were observed. The growth curve of the cells was plotted by CCK-8 method, the expression of CD29,CD45,CD90 was detected by immunofluorescence, the differentiation was induced by fat-forming and osteogenic medium, and the differentiation of cells was identified by oil red Oand alizarin red staining. Results the morphology of ADSC isolated from the adipose tissue of cryopreserved SD rats was long fusiform, with a good proliferative ability and a typical "S" type of growth curve, and the strong expression of CD29,CD90,CD45 in the cells was confirmed by immunofluorescence technique. Oil red O staining was positive after adipogenic induction and alizarin red staining was positive after osteogenesis induction. Conclusion ADSC. can be isolated from adipose tissue of cryopreserved SD rats.
【作者单位】：中国人民解放军武汉总医院医学实验科;
【分类号】：R329.2

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