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

描述(申请人提供)：本研究的长期目标是开发从组织学上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的结肠隐窝的方法。在培养中保持的分离的结肠隐窝将提供一个独特的培养系统，用于在临床前环境中检查潜在的化学预防药物的效果，以及检查结肠生长调节的各个方面。同样的培养系统有望被证明有助于从正常和异常的隐窝中分离和鉴定干细胞。为了实现这一建议的目标，我们将做以下工作：特定的目标：从组织学上正常的人结肠组织和腺瘤性息肉中分离隐窝；开发保持隐窝组织结构和生化功能的培养条件，允许组织学正常的隐窝通过增殖、分化和凋亡的方式在空间和时间上适当地进展，同时允许来自癌前病变息肉的组织继续生长和扩张。

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.