Isolation and in vitro maintenance of intact colonic crypts from normal human col

摘要本研究的长期目标是开发从组织学正常的人结肠组织和腺瘤性息肉/恶性肿瘤中分离和体外维持完整结肠隐窝的方法。在培养中保持的分离结肠隐窝将为在临床前环境中检查潜在化学预防剂的效果以及检查结肠生长调节的各个方面提供独特的培养系统。同样的培养系统将有望被证明对从正常和异常隐窝中分离和表征干细胞有用。具体目标：从组织学正常的人类结肠组织和腺瘤性息肉中分离隐窝；培养维持隐窝组织结构和生化功能的培养条件，允许组织正常的隐窝在适当的空间和时间内通过增殖、分化和凋亡进行进展，同时允许来自癌前腺瘤性息肉的组织持续生长和扩张。

DESCRIPTION (provided by applicant): Abstract the long-term goal of the research proposed here is to develop methods for the isolation and in vitro maintenance of intact colonic crypts from histologically normal human colon tissue and from adenomatous polyps / malignant tumors. The isolated colonic crypts maintained in culture will provide a unique culture system for examining the effects of potential chemopreventive agents in a preclinical setting as well as for examining various aspects of growth regulation in the colon. The same culture system will, hopefully, prove useful for isolation and characterization of stem cells from normal and aberrant crypts. To achieve the goals of this proposal, we will do the following: Specific Aim: Isolate crypts from histologically normal human colon tissue and from adenomatous polyps; develop culture conditions that maintain crypt histological structure and biochemical function and that allow for progression of the histologically-normal crypts through proliferation, differentiation an apoptosis in a spatially and temporally appropriate manner, while allowing for continued growth and expansion of the tissue derived from premalignant adenomatous polyps.