期刊
STEM CELL REPORTS
卷 10, 期 1, 页码 314-328出版社
CELL PRESS
DOI: 10.1016/j.stemcr.2017.11.004
关键词
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资金
- Japan Tobacco
- Ministry of Education, Science, Sports and Technology of Japan [25860353, 23229004]
- Japan Agency for Medical Research and Development (AMED)
- Grants-in-Aid for Scientific Research [25860353] Funding Source: KAKEN
Gut epithelial organoids are routinely used to investigate intestinal biology; however, current culture methods are not amenable to genetic manipulation, and it is difficult to generate sufficient numbers for high-throughput studies. Here, we present an improved culture system of human induced pluripotent stem cell (iPSC)-derived intestinal organoids involving four methodological advances. (1) We adopted a lentiviral vector to readily establish and optimize conditioned medium for human intestinal organoid culture. (2) We obtained intestinal organoids from human iPSCs more efficiently by supplementing WNT3A and fibroblast growth factor 2 to induce differentiation into definitive endoderm. (3) Using 2D culture, followed by re-establishment of organoids, we achieved an efficient transduction of exogenous genes in organoids. (4) We investigated suspension organoid culture without scaffolds for easier harvesting and assays. These techniques enable us to develop, maintain, and expand intestinal organoids readily and quickly at low cost, facilitating high-throughput screening of pathogenic factors and candidate treatments for gastrointestinal diseases.
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