脂肪来源基质细胞的扩增及多分化潜能研究

Amplification and multi-lineage differentiation potential of adipose derived stromal cells

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100ml脂肪抽吸物中可分离得到2×10^7-4×10^7个有核细胞,60d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。

Objective To establish the method of the separation and amplification in vitro of human adipose derived stromal ceils （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 ceil time course and cumulative population doubling. ADSCs were induced into adipocytes and neuron-like ceils by different factors to assess the committed differentiations. Results 2 × 10^7 -4 × 10^7 nucleated cells were obtained from 100 ml lipoaspirate. ADSCs were stably proliferated and transferred in vitro till P8. ADSCs differentiated into adipose ceils and neuron like ceils certified by Oil red O staining, immunocytochemical stain and RT-PCR, Conclusions The method for ampliilcation and culture of ADSCs is established, the amplificated and cultured ADSCs have vigorous proliferation activity and potential to differentiate into adipocytes and neuron-like ceils, which can possibly be served as a promising mature muhipotent stem ceil for tissue engineering and ceil therapy.