發(fā)布時(shí)間：2018-08-22 14:32

【摘要】：目的制備人脂肪脫細(xì)胞基質(zhì)并初步評(píng)價(jià)其細(xì)胞相容性,為構(gòu)建組織工程脂肪尋找理想的支架材料。方法以接受乳腺癌根治術(shù)或乳腺癌改良根治術(shù)患者自愿捐贈(zèng)的腹壁皮下脂肪組織作為實(shí)驗(yàn)標(biāo)本,取其中一部分經(jīng)反復(fù)凍融、有機(jī)溶劑抽提以及酶消化處理制備脂肪脫細(xì)胞基質(zhì),大體觀察其性狀,行組織學(xué)、免疫組織化學(xué)、DAPI熒光染色及掃描電鏡觀察材料脫細(xì)胞效果、重要組成成分(Ⅳ型膠原、層粘連蛋白、纖連蛋白)保留情況及微觀結(jié)構(gòu)。取另一部分脂肪組織采用酶消化法分離培養(yǎng)人脂肪干細(xì)胞(human adipose-derived stem cells,hADSCs),取第3代細(xì)胞分別與脂肪脫細(xì)胞基質(zhì)及濃度為100%、75%、50%、25%的浸提液復(fù)合培養(yǎng)。采用MTT法檢測(cè)細(xì)胞毒性,掃描電鏡觀察細(xì)胞黏附情況,活/死細(xì)胞染色評(píng)價(jià)細(xì)胞增殖情況。結(jié)果脂肪脫細(xì)胞基質(zhì)基本保持原組織外形;無細(xì)胞殘留且保留較完整細(xì)胞外基質(zhì)成分,含豐富的Ⅰ型膠原,無DNA、脂質(zhì)殘留;其以膠原纖維為主,含豐富的層粘連蛋白和纖連蛋白。脂肪脫細(xì)胞基質(zhì)不同濃度浸提液與細(xì)胞復(fù)合培養(yǎng)1、3、5 d,細(xì)胞毒性均為0~1級(jí)。hADSCs在脂肪脫細(xì)胞基質(zhì)中黏附、增殖良好。結(jié)論利用反復(fù)凍融、有機(jī)溶劑抽提以及酶處理法所制備的脂肪脫細(xì)胞基質(zhì)保留了較完整的細(xì)胞外基質(zhì)成分,細(xì)胞相容性好,有望成為組織工程脂肪支架材料。
[Abstract]:Objective to prepare human adipose acellular matrix and evaluate its cytocompatibility to find an ideal scaffold for the construction of tissue engineered fat. Methods the abdominal subcutaneous adipose tissue from patients undergoing radical mastectomy or modified radical mastectomy was used as experimental specimen, and some of them were prepared by repeated freeze-thaw, organic solvent extraction and enzyme digestion to prepare adipose acellular matrix. The characteristics, histology, DAPI fluorescence staining and scanning electron microscope were used to observe the acellular effect, the retention and microstructure of important components (type 鈪，

本文編號(hào)：2197383