利用ADSCs在生物反应器中构建小口径血管的初步研究

Preliminary Study on Small Blood Vessel Constructed in the Bioreactor Using ADSCs

目的尝试利用人的脂肪基质干细胞(Adipose-derived stromal cell,ADSCs)在生物反应器中构建组织工程化小血管。方法采用脂肪抽吸术获取人脂肪组织,将脂肪细胞分离、传代培养;对培养的第三代细胞进行脂肪干细胞表型鉴定;同时称取一定量的聚羟基乙酸(PGA)制成片状材料,消毒晾干备用,将细胞接种在PGA材料上,静态培养半天后置于生物反应器中动态培养4周,然后进行初步检测。结果从形态学观察,细胞伸展呈成纤维细胞样,生长有方向性,5～6d后可见克隆开始形成。对第三代ADSCs进行检测,结果显示除CD166、CD105有高的表达外,CD14、CD34表达很低。从反应器取出的样品能形成血管样组织,扫描电镜观察:样品基质分泌旺盛。结论采用组织工程方法,从动态培养的角度可以在体外利用ADSCs构建血管形状的组织,为进一步研究ADSCs在血管组织工程中的应用提供了一种好的方法。

Objectivs Construct tissue engineered small-caliber blood vessel in bioreactor using Adipose-derived stem cells （ADSCs）. Methods ADSCs were isolated and subeultured from adipose tissue of patients who received suction lipeetomy. The phenotypieal analysis of the 3rd generation of ADSCs was performed. After constructed with ADSCs and PGA flakes, the cell-material compound was co-cultured in static condition and placed in a dynamic culture bioreaetor for future 4 weeks,followed by preliminary analysis. Results During monolayer expansion, the cells appeared fibroblasts-like morphology with numerous clones formation Flow eytometry analysis showed that the third generation of ADSCs highly expressed CD166 and CD105, while little CD14 and CD34. Blood-vessel-shaped tissues were formed in the bioreaetor,mueh extraeellular matrix was observed by scanning electron microscopy. Conclusion Blood vessel shaped tissue can be constructed in vitro using ADSCs under a dynamic culture condition,establishment of this system may become an effective tool for further investigation and broaden the application of ADSCs in the blood vessel tissue engineering.