脂肪干细胞与三维支架体内外培养构建的组织工程化气管

背景：气管替代物包括自体组织皮瓣、气管同种异体移植、人工材料支架和无活力组织移植等,但均因为相关严重并发症和获取困难等因素使临床应用遇到很大困难。目的：利用脂肪干细胞与聚乳酸-乙醇酸共聚物（poly-D,L-lactio-co-glycolicacid,PLGA）、聚三亚甲基碳酸酯（poly（trimethylenecarbonate）,PTMC）共聚物支架构建组织工程化气管支架模型。方法：组织块法原代分离培养SD大鼠脂肪干细胞,脂肪干细胞行流式细胞术及多向分化能力鉴定,分别种植于PLGA-PTMC支架,经体内体外培养后行免疫组织化学及扫描电镜观察。结果与结论：大鼠脂肪干细胞接种于支架后,呈球形,伸展出伪足,均匀贴附PLGA-PTMC支架,细胞间相互融合成团;经体内培养后,新生毛细血管丰富。PLGA-PTMC支架具有良好的生物相容性,无细胞毒性,其多孔的三维立体状结构适合脂肪干细胞黏附生长。经体内、体外培养得到组织工程化气管模型,新生血管丰富,可以作为有效的气管替代物。

Construction of tissue engineering trachea by in vitro and in vivo culture of adipose-derived stem cells in three-dimensional scaffolds

BACKGROUND：Trachea substitutes include autologous tissue flap,allogeneic tracheal transplant,artificial scaffolds and non-dynamic tissue transplant.However,they are hardly used in the clinic because of severe complications and difficult harvest.OBJECTIVE：To establish the tissue-engineered tracheal scaffold model by adipose-derived stem cells,poly-D,L-lactio-co-glycolic acid and poly（trimethylenecarbonate）.METHODS：Adipose-derived stem cells of Sprague Dawley rats were isolated and cultured by tissue block method.Cell surface markers were evaluated by flow cytometry and multilineage differentiation,and the cells were implanted on the poly-D,L-lactio-co-glycolic-poly（trimethylenecarbonate） scaffold.After in vitro and in vivo culture,the adipose-derived stem cells were observed by immunohistochemistry and scanning electron microscope.RESULT AND CONCLUSION：Adipose-derived stem cells were spherical and well attached to the scaffolds with lamellipodia and conglobate connection after inoculated on the poly-D,L-lactio-co-glycolic-poly（trimethylenecarbonate） scaffold.After in vitro culture,the newborn capillary could be seen.The poly-D,L-lactio-co-glycolic-poly（trimethylenecarbonate） scaffold scaffolds with porous structure were highly biocompatible and non-cytotoxic as determined by adipose-derived stem cells adhesion and proliferation.Neotrachea fabricated with combined in vitro and in vivo culture system using poly-D,L-lactio-co-glycolic-poly（trimethylenecarbonate） scaffold and adipose-derived stem cells can meet the requirement of tracheal replacement,this model serve as a new way to engineer a vascularized trachea.