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目的:建立大肠癌癌旁组织体外3-D培养实验平台,观察距瘤体不同距离肠上皮组织体外生长特性。方法:大肠癌手术时,分别从肿瘤近端距瘤体2cm、5cm、10cm的部位取肠上皮组织,运用重复贴壁法进行3-D培养,观察组织生长形态特征、肠隐窝微架构变化特征。结果:组织完全贴壁,上皮粘膜位于“气-液”交界进行培养有利于组织块的存活与成长;培养液中牛血清浓度对组织块生长及肠隐窝结构的维持有很大的影响,牛血清的存在不利于组织的存活,其浓度越高组织生长越受到抑制;血清的存在也很不利于隐窝微架构的存在,血清浓度越高肠隐窝微架构丧失的越快。结论:无血清培养条件下,结肠癌旁近、中、远三个不同部位的组织都可以进行体外培养,但培养过程中,维持隐窝最初离体时的结构还是比较困难,培养3d隐窝结构变化不大,7-21d隐窝结构内细胞会逐渐降解。血清的存在不利于隐窝的结构维持,隐窝结构丧失得更快。
OBJECTIVE: To establish an in vitro 3-D culture experimental platform of colorectal cancer adjacent tissues and observe the growth characteristics of intestinal epithelial tissue in vitro at different distances from the tumor. Methods: The intestine tissues of 2 cm, 5 cm and 10 cm from the tumor proximal to the tumor were collected during operation of colorectal cancer. The 3-D culture was performed by repeated adherent method. The morphological changes of the tissue, the micro-structure of intestinal crypt feature. Results: Tissue completely adherent, epithelial mucosa located at the junction of “gas-liquid” was conducive to the survival and growth of tissue mass; the concentration of bovine serum in culture medium had significant effect on the growth of tissue mass and the structure of intestinal crypt The existence of bovine serum is not conducive to the survival of the tissue, the higher the concentration of tissue growth inhibition; the presence of serum is also not conducive to the existence of crypt micro-architecture, the higher the serum concentration, the loss of intestinal crypt micro-architecture faster. CONCLUSION: Under the condition of serum-free culture, the tissues in the proximal, middle, and distal colon of colon cancer can be cultured in vitro. However, it is still difficult to maintain the structure of crypt initially in vitro during culture. Structure changes little, 7-21d crypt cells within the gradual degradation. The existence of serum is not conducive to maintaining the structure of the crypt, crypt structure loss faster.