类有机物
细胞外基质
细胞生物学
基质(化学分析)
结直肠癌
细胞培养
化学
生物
癌症
遗传学
色谱法
作者
Tao Tan,Yumiko Hirokawa,Jordan Clarke,Anuratha Sakthianandeswaren,Oliver M. Sieber
出处
期刊:Bio-protocol
[Bio-Protocol]
日期:2022-01-01
卷期号:12 (8)
被引量:6
标识
DOI:10.21769/bioprotoc.4394
摘要
Three-dimensional culture of human normal colorectal epithelium and cancer tissue as organoids and tumoroids has transformed the study of diseases of the large intestine. A widely used strategy for generating patient-derived colorectal organoids and tumoroids involves embedding cells in domes of extracellular matrix (ECM). Despite its success, dome culture is not ideal for scalable expansion, experimentation, and high-throughput screening applications. Our group has developed a protocol for growing patient-derived colorectal organoids and tumoroids in low-viscosity matrix (LVM) suspension culture. Instead of embedding colonic crypts or tumor fragments in solid ECM, these are grown suspended in medium containing only a low percentage of ECM. Compared with dome cultures, LVM suspension culture reduces the labor and cost of establishing and passaging organoids and tumoroids, enables rapid expansion, and is readily adaptable for high-throughput screening. Graphic abstract: Generation of organoids and tumoroids from human large intestine using LVM suspension culture (Created with BioRender.com).
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