明胶微冰胶支架有利于脂肪来源间充质干细胞体外干性维持并提高其体内应用价值  

Gelatin microcryogels benefit human mesenchymal stem cells derived from adipose tissue to maintain stemness in vitro and promote application value in vivo

作　　者：毛晓晶 曾洋[2] 韩钦[1] 赵春华[1] 

机构地区：[1]中国医学科学院基础医学研究所组织工程中心,北京100005 [2]清华大学医学院生物医学工程,北京100084

出　　处：《基础医学与临床》2015年第5期610-614,共5页Basic and Clinical Medicine

基　　金：国家重大科学研究计划(2011CB964901)

摘　　要：目的研究人脂肪来源间充质干细胞(ADSCs)与明胶微冰胶材料体外联合培养时,材料是否能维持间充质干细胞的生物学特性。方法 ADSCs种植于明胶微冰胶材料后进行Calcein-AM和PI活细胞染色检测细胞活力,细胞滴度蓝法检测细胞增殖能力,定量PCR检测干性基因OCT4、Nanog、SOX2表达情况,以及在成脂成骨诱导过程比较ADSCs在二维环境和种植于明胶微冰胶材料后的分化潜能。结果 ADSCs在三维明胶微冰胶支架材料中能保持较高活性,增殖能力不受影响,干性基因表达上调,成脂成骨分化相关基因表达水平比二维诱导环境低。结论明胶微冰胶可以为ADSCs提供一个较二维培养更好的微环境,有利于ADSCs体外干性维持,从而在干细胞移植法治疗相关疾病时以非侵入性的细胞传递方式发挥更长期的应用价值。Objective To investigate whether the gelatin microcryogels can maintain biological characteristics of human mesenchymal stem cells from adipose tissue（ ADSCs） and enhance the stemness of ADSCs when they are cocultured in vitro. Methods Calcein-AM and PI staining were conducted to test cell viability of ADSCs plated in gelatin microcryogels. Cell titer-blue assay was used to examine the cellular proliferating capacity. Real-time PCR was used to evaluate the expression level of the stemness genes OCT4,Nanog,SOX2. Adipogenic and osteogenic differentiation were induced and compared in ADSCs plated in gelatin microcryogels and in conventional environment. Results ADSCs were biologically active in the 3D scaffolds of gelatin microcryogels. Proliferation rate was not influenced. Stemness genes expression was up-regulated. ADSCs plated in gelatin microcryogels still had osteogenic and adipogenic differentiation ability,but the related genes expression levels were lower in comparison with ADSCs in conventional inducing condition. Conclusions Gelatin microcryogels can provide proper microenvironmental factors for ADSCs stemness maintenance,so as to exert the long-term application value in the diseases treatedwith stem cell by a minimally-invasive delivery method.

关 键 词：人脂肪间充质干细胞 明胶微冰胶 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]