人脂肪干细胞的分离培养与鉴定

Isolation, culture and identification of human adipose-derived stem cells

目的探索一种体外分离培养人脂肪干细胞(h ADSC)的有效方法。方法选择健康女性5例,年龄25~45岁;无任何其他系统性疾病。对其应用抽脂术获取人脂肪组织,并用Ⅰ型胶原酶进行消化,贴壁培养,传代扩增,绘制不同代数细胞增殖曲线,诱导hADSC向成骨、成脂细胞分化,确定其分化潜能,并分别用茜素红和油红O染色进行鉴定。结果原代hADSC第2天开始贴壁,形态呈纺锤型,类似纤维细胞,增殖较快,呈漩涡状排列。第3代至第5代hADSC生长曲线类似,都呈S形;成骨、成脂诱导2周后进行茜素红染色和油红O染色,结果都为阳性。结论通过对抽脂术获得的人脂肪组织进行分离培养,可以得到有效的hADSC,且具有向成骨、成脂细胞方向分化的能力,可作为进一步实验研究的种子细胞。

Objective To explore an effective method for isolating and identifying human adipose-derived stem cells（hADSC ）.Methods A total of 5 healthy females, aged 25-45 years old without systemic disease were enrolled. The liposuction was used to obtain human adipose tissue, and collagenase type Ⅰ digestion was adopted to isolate and culture. Morphology and cell growth curve were characterized, then hADSC was inducted to osteogenic and adipogenic differentiation, and identified by alizarin red and oil red O staining, respectively. Results The primary hADSC adhered to wall at 2nd days, the adherent cells were fusiform-shaped as fibroblasts, and proliferated faster with whirlpool. The growth curves of hADSC form passages 3 to 5were similar with S-shape. The results of alizarin red staining and oil red O staining after osteogenic, adipogenic induction for2 weeks were positive. Conclusion The hADSC can be obtained from human adipose tissue from liposuction by isolation and culture. With the potential to differentiate into osteogenic and adipogenic cells, hADSC can be used as seed cells in further experimental study.