经RNAi诱导前脂肪细胞构建组织工程化脂肪组织.doc

经RNAi诱导前脂肪细胞构建组织工程化脂肪组织[摘要]目的：应用基因工程和组织工程的原理和方法,通过改造人前体脂肪细胞，提高其活性后，于裸鼠背部皮下构建组织工程化脂肪组织。方法：采用原代消化细胞培养法培养出前脂肪细胞,利用RNAi技术，诱导前脂肪细胞内的一个重要凋亡调控基因Bax基因表达沉默,然后接种到聚乳酸-乙醇酸共聚物(PLGA)支架上,种植在20只雌性裸鼠皮下,对获得的脂肪组织进行组织学评价。结果：经过改造后的前脂肪细胞活性良好,术后裸鼠全部成活，前脂肪细胞能够在支架上良好的存活，生长和增殖。结论：经过改造后的前脂肪细胞具有良好的活性，并且PLGA- 前脂肪细胞复合体在裸鼠体内可形成脂肪细胞,值得进一步研究, 以探索治疗软组织缺损的新途径。 [关键词]RNAi；Bax基因；组织工程；脂肪组织 [中图分类号]Q813[文献标识码]A [文章编号]1008-6455（2008）10-1479-04 RNAi induced preadipocytes to build tissue engineering adipose tissue ZHUANG Jian-hua1，ZHUO Yang2，ZENG Xing-ye2 (1.Department of Surgery, Lucheng People’s Hospital, Wenzhou 325000,Zhejiang,China; 2.Department of Surgery, the 118th Hospital of PLA) ObjectiveUsing genetic engineering and tissue engineering principles and methods, through the transformation of preadipocytes, enhancing its activity,and to construct tissue engineering adipose tissue in the back skin of nude mice.Methods Using primary-digest method to culture preadipocytes, using RNAi technology to induct an important apoptosis gene Bax gene expression silence in preadipocytes，and then inoculated into polylactic acid - glycolic acid copolymer (PLGA) scaffold , planted in 20 female nude mice, the adipose tissue was carried out histological evaluation. Results After ransformation, the activity of preadipocytes is good, after all the mice survived. preadipocytes can survive well in the scaffold, growing and proliferation.ConclusionsAfter ransformation, preadipocytes has good activity, and PLGA- preadipocytes can be formed fat cells in nude mice, it is worth further study to explore the treatment of soft tissue defects in new ways. Key words：RNAi；Bax gene；tissue engineering；adipose tissue 自体脂肪组织是修复各类组织缺损的最佳填充物，但体内脂肪组织一旦离开活体常易出现变性、坏死，如何获取活性脂肪细胞，是脂肪组织移植成功的关键。前脂肪细胞是一类能够增殖并向脂肪细胞分化的特异性前体细胞，为构建组织工程化脂肪组织的理想种子细胞。本研究从人体脂肪组织中提取人前脂肪细胞作为种子细胞，经体外培养扩增后，利用RNAi技术，诱导前脂肪细胞内的一个重要的凋亡调控基因Bax基因表达沉默,以促进前脂肪细胞的增殖和分化，抑制其凋亡，经上述处理的前脂肪细胞进行相关检测后，再和可降解人工细胞外基质（PLGA）混合培养，于裸鼠背部皮下体内构建组织工程化脂肪组织，以探索治疗软组织缺损的新的途径。 1材料和方