人脂肪来源干细胞部分生物学性状的实验研究

Experimental study for some biological characteristics of human adipose-derived stem cells

目的对体外分离培养人脂肪来源干细胞的部分生物学性状进行分析。方法将肿胀吸脂术获得的脂肪组织通过酶消化法分离培养脂肪来源干细胞，采用流式细胞仪检测细胞的表面标记；取第2代细胞进行成脂、成内皮细胞诱导。结果体外培养的原代脂肪来源干细胞成分较复杂，形态各异，细胞生长缓慢；细胞传至第2代形态渐趋一致，以梭形为主，多次传代后细胞仍保持较强增殖能力。原代细胞CD29、CD31、CD34、CD49d、CDl05、CDl66表达为阳性；第2代细胞CD31、CIM9d转为阴性，而CD29、CD105、CD166、Stro-1、Flk-1表达比例显著多于原代细胞。成脂诱导后细胞内可见大量脂滴形成，油红O染色阳性；成内皮细胞诱导后，细胞表达CD31、vWF等成熟内皮细胞标志。结论通过吸脂术获得的大量原代脂肪干细胞是由极其复杂的各种细胞集合而成，在体外易于分离培养和传代扩增，传代后的细胞在特定条件下可诱导分化为脂肪细胞和血管内皮细胞，并表达间充质干细胞相关的表型，是组织工程理想的种子细胞。

Objective To investigate the biological characteristics of adipose-derived stem cells （ADSCs） isolated and cultured in vitro. Methods Through isolating culture by enzymatic digestion, ADSCs were harvested from adipose tissue, which was acquired by tLunescent liposuction, and then the surface marker were detected by flow cytometry. Adipogenic differentiation and endothelial differentiation of ADSCs on 2nd generation was induced respectively. Results The component of the primary ADSCs was complicated with vary morphous and slow growth, the cellular morphous on the 2nd generation became identical gradually with a spindle-shaped appearance mainly. The proliferation activity kept stronger after high passage. The expression of CD29, CD31, CD34, CD49d, CD105 and CD166 at primary ADSCs were positive , however, the expression of CD31 and CIM9d turned into negative on the secondary ADSCs, while the expression of CD29, CD105, CD166, Stro-1 and Flk-I were stronger than those on the primary ADSCs. ADSCs after adipogenic induction demonstrated numerous intracellular lipid droplets by Oil red O staining. The expression of CD31 and vWF, the markers of mature endothelial cells, were positive after endothelial induction. Conclusion The numerous primary ADSCs isolated from human liposuction aspirates are composed of many different stem cells, which are easy to be isolated and have strong proliferative ability. ADSCs also can be induced to differentiate into adipose cells and endothelial cells by the action of the induction agent. So ADSCs are a kind of ideal seed cells for tissue engineering which express cell surface markers of stem cells.