猪小肠黏膜下层基质支架材料复合脂肪基质干细胞的生物相容性

背景：小肠黏膜下层具有良好的细胞、组织相容性和降解性，是一种理想的组织工程支架材料，将脂肪基质干细胞与小肠黏膜下层复合后进行定向诱导，可构建靶组织，具有一定的临床应用潜能。 目的：制备脱细胞猪小肠黏膜下层基质，并检验其与家兔脂肪基质干细胞的生物相容性。 方法：使用酶消化-高盐水脱细胞法处理猪小肠黏膜下层，石蜡切片检测脱细胞效果，扫描电镜观察小肠黏膜下层表面结构。体外分离培养家兔脂肪基质干细胞，分别将第3代脂肪基质干细胞单面复合和双面复合至小肠黏膜下层培养1周观察材料上下表面细胞复合情况。 结果与结论：小肠黏膜下层材料为白色半透明膜状物，石蜡切片显示细胞去除彻底，扫描电镜显示小肠黏膜下层黏膜结构紧密，浆膜面纤维结构松散。脂肪基质干细胞与材料复合后，通过相差显微镜观察到细胞在小肠黏膜下层上附着生长，扫描电镜检测提示单面复合脂肪基质干细胞的小肠黏膜下层，仅在上表面有大量细胞生长，下表面无细胞或仅有少量细胞生长，双面复合脂肪基质干细胞的小肠黏膜下层上下表面均可见大量细胞融合生长，石蜡切片可见细胞贴附于材料表面生长。提示酶消化-高渗盐水法可彻底去除小肠黏膜下层表面细胞成分，小肠黏膜下层对脂肪基质干细胞的生长具有良好的支持作用。 关键词：小肠黏膜下层；酶消化-高渗盐水脱细胞法；脂肪基质干细胞；生物相容性；支架材料 doi:10.3969/j.issn.1673-8225.2010.03.009

Biocompatibility of porcine small intestinal submucosa and adipose derived mesenchymal stem cells

BACKGROUND: Small intestinal submucosa (SIS) has good compatibility with cells and tissues, and has good degradability. It is an ideal scaffold for tissue engineering. Inducing adipose derived mesenchymal stem cells (ADSCs) seeded on SIS can construct target tissues, which has the potential to be used in clinical treatment. OBJECTIVE: To prepare decellularized porcine SIS matrix, and testify its biocompatibility with rabbit ADSCs cultured in vitro. METHODS: SIS was processed by enzyme digestion-hypertonic saline decellularization, lyophilized at low temperature, and sterilized by gamma radiation. Paraffin sections were used to observe the effect of decellularization of SIS, and the surface structures of SIS were observed by scanning electron microscope (SEM). Rabbit ADSCs were isolated and cultured, and passage 3 ADSCs were seeded onto one side or both sides of SIS. After one week of co-culture, the cell-scaffold composites were observed. RESULTS AND CONCLUSION: SIS was white and semi-transparent film. Paraffin sections showed no cells on SIS matrix; electron microscopy showed loose weave structure of serosal surface and dense packing structure of mucosal surface. After one week of co-cultivation, plenty of ADSCs were observed on the surface of SIS. In ADSCs seeded onto one side of SIS group, a large number of cells grew on the superior surface, and few even no cells were observed on inferior surface of SIS. When ADSCs were seeded onto both sides of SIS, cells adhered to SIS in paraffin sections. Results show that enzyme digestion-hypertonic saline decellulariation can decellularize SIS completely, and SIS can support ADSCs growth.