【2017年整理】脂肪与来源基质细胞的扩增及多分化潜能研究.docVIP

【摘要】 目的 建立人脂肪来源的基质细胞(ADSCs)的分离和扩增方法，探讨ADSCs的多向分化潜能。方法 采用胶原酶消化法分离扩增人ADSCs,测定其生长的经时变化、累积生长倍数等；应用不同因子诱导ADSCs向脂肪细胞及神经元样细胞分化并进行鉴定。结果 每100 ml脂肪抽吸物中可分离得到2×107～4×107个有核细胞，60 d内扩增至P8；油红O染色、免疫细胞化学染色及RTPCR证实ADSCs经诱导可分化为脂肪细胞及神经元样细胞。结论 建立了ADSCs的扩增培养方法，扩增培养的ADSCs具有旺盛的增殖能力，并且具有向脂肪细胞及神经元样细胞分化的潜能。有望作为一种成体多能干细胞应用于组织工程及细胞治疗。 【关键词】 脂肪来源基质细胞；多分化潜能；脂肪细胞；神经元样细胞 　　Amplification and multilineage differentiation potential of adipose derived stromal cells 　　WANG WenJia, JIAO Ping, XU TianMin, et al 　　Institute of Frontier Medical Science, Jilin University, Changchun 130021, Jilin, China 　　【Abstract】 Objective To establish the method of the separation and amplification in vitro of human adipose derived stromal cells (ADSCs) and to investigate its multilineage differentiation potential.Methods ADSCs were isolated and amplificated by collagenase digestion from lipoaspirate to detect the proliferation kinetics of ADSCs through cell time course and cumulative population doubling. ADSCs were induced into adipocytes and neuronlike cells by different factors to assess the committeddifferentiations.Results 2×107～4×107 nucleated cells were obtained from 100 ml lipoaspirate. ADSCs were stably proliferated and transferred in vitro till P8. ADSCs differentiated into adipose cells and neuron like cells certified by Oil red O staining, immunocytochemical stain and RTPCR. Conclusions The method for amplification and culture of ADSCs is established, the amplificated and cultured ADSCs have vigorous proliferation activity and potential to differentiate into adipocytes and neuronlike cells, which can possibly be served as a promising mature multipotent stem cell for tissue engineering and cell therapy. 　　【Key words】Adipose derived stromal cells (ADSCs); Multilineage differentiation potential; Adipocytes; Neuronlike cells 骨髓基质细胞（bone marrow stromal cells，BMSCs）是较早被认识的成体干细胞之一，在细胞治疗、基因治疗及再生医学领域具有重要的临床应用价值。但骨髓的抽取过程患者不易接受，而且在1×106个骨髓单个核细胞中仅有2～5个BMSCs，使大量获取BMSCs受到限制。脂肪来源的基质细胞（adipose deriv