脂肪脱细胞基质的制备和初步评价
发布时间:2018-08-22 14:32
【摘要】:目的制备人脂肪脱细胞基质并初步评价其细胞相容性,为构建组织工程脂肪寻找理想的支架材料。方法以接受乳腺癌根治术或乳腺癌改良根治术患者自愿捐赠的腹壁皮下脂肪组织作为实验标本,取其中一部分经反复冻融、有机溶剂抽提以及酶消化处理制备脂肪脱细胞基质,大体观察其性状,行组织学、免疫组织化学、DAPI荧光染色及扫描电镜观察材料脱细胞效果、重要组成成分(Ⅳ型胶原、层粘连蛋白、纤连蛋白)保留情况及微观结构。取另一部分脂肪组织采用酶消化法分离培养人脂肪干细胞(human adipose-derived stem cells,hADSCs),取第3代细胞分别与脂肪脱细胞基质及浓度为100%、75%、50%、25%的浸提液复合培养。采用MTT法检测细胞毒性,扫描电镜观察细胞黏附情况,活/死细胞染色评价细胞增殖情况。结果脂肪脱细胞基质基本保持原组织外形;无细胞残留且保留较完整细胞外基质成分,含丰富的Ⅰ型胶原,无DNA、脂质残留;其以胶原纤维为主,含丰富的层粘连蛋白和纤连蛋白。脂肪脱细胞基质不同浓度浸提液与细胞复合培养1、3、5 d,细胞毒性均为0~1级。hADSCs在脂肪脱细胞基质中黏附、增殖良好。结论利用反复冻融、有机溶剂抽提以及酶处理法所制备的脂肪脱细胞基质保留了较完整的细胞外基质成分,细胞相容性好,有望成为组织工程脂肪支架材料。
[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,
本文编号:2197383
[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,
本文编号:2197383
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