猪角朊细胞复合小肠黏膜下层体外培养实验

目的 探讨脱细胞小肠黏膜下层（SIS）作为组织工程皮肤支架的可行性.方法 将猪第二代角朊细胞接种于SIS上,接种后第11、13、15、17天通过直接染色、石蜡组织切片、HE染色、抗层粘连蛋白免疫组化染色、透射电镜等方法,观察角朊细胞在SIS上的生长情况.结果 猪角朊细胞接种在SIS上11d形态良好,部分区域形成2～3层细胞组成的复层结构,免疫组织化学染色显示在角朊细胞与SIS之间的细胞外基质中抗层粘连蛋白呈连续阳性表达.13、15、17 d细胞复层结构增多,细胞之间连接紧密,透射电镜下可见细胞内有张力微丝,细胞间有桥粒连接,角朊细胞与SIS之间可见基底膜形成.结论 在猪角朊细胞体外培养中,SIS是良好的细胞黏附支架.

The experiment of porcine keratinocytes cultured on porcine small intestinal submucosa in vitro

OBJECTIVE: To investigate the feasibility of acellular porcine small intestinal submucosa (SIS as bioscaffold of tissue engineering skin. METHODS: The second passage keratinocytes were seeded on SIS, after seeded for 11, 13, 15, 17 days, the keratinocytes/SIS composites were observed by dye directly, histopathology, immunohistochemical studies with monoclonal antibodies against laminin and transmission electron microscope (TEM). RESULTS: At eleventh day, keratinocytes were growth very well on the surface of SIS, there are 2-3 cell layers on partial of the SIS surface, the continued expression of laminin can be detected between the keratinocytes and the surface of SIS. After 13, 15, 17 days this stratified structure increased, cells contact more closely, the tonofibirls in cells, desmosome between cells and the basal membrane between the keratinocytes and the surface of SIS can be seen with TEM. CONCLUSIONS: SIS is a kind of good bioscafold in the culture of porcine keratinoytes in vitro.