改良法分离大鼠IOUs构建小肠的初步研究

目的采用改良方法分离大鼠小肠上皮类器官单位(IOUs),鉴定IOUs裸鼠体内发育能力。方法取出生5～8d大鼠的小肠组织,采用改良分离法获得IOUs,与细胞外基质胶混合后植入裸鼠皮下,定期取材,组织学鉴定。结果采用改良方法分离时间明显缩短,体内植入所有实验组均有多个囊泡样组织形成,植入2周后部分囊腔凹陷可见隐窝样结构,2周后部分囊泡内壁可见分化较成熟的上皮细胞突出,类似绒毛结构;上皮下可见少量平滑肌样细胞围绕囊状管腔周围。结论初步证实改良方法分离出IOUs用于组织工程肠管构建的可行性,并为出生后组织修复重构的研究奠定了基础。

Preliminary Study on Reconstructing Rat Intestine with Intestinal Epithelial Organoid Units Isolated by Modified Method

Objective To isolate the rat intestinal epithelial organoid units(IOUs) by modified methods and to identify the viability of IOUs in vivo of nude mouse. Methods IOUs were recovered from 5 to 8-day-old Wistar rat pups by modified isolation method, mixed with ECM gel, and infused subcutaneously into nude mice. They were sacrificed from 1 to 4 weeks after implantation at weekly intervals. Identification with histological and immunohistochemical stain were performed. Results With the application of new isolation methods, the dissociated time shortened distinctly. Every harvested graft formed cyst-like tissue. The lumina of cysts were full of mucus and debris of apoptosis cells. Intestinal villus and crypts could be perceived by histological stain in the graft harvested two weeks after implantation. There were also smooth muscle-like cells and positive cells of PGP9.5 immunohistochemical stain dispersing around the intestinal epithelium. Conclusion IOUs isolated with modified methods could serve as seed cells for reconstructing rat intestine and a model for other research fields.