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Merck

Introducing the Stem Cell ASCL2 Reporter STAR into Intestinal Organoids.

STAR protocols (2020-12-31)
Maria C Heinz, Koen C Oost, Hugo J G Snippert
要旨

Patient-derived organoids maintain functional and phenotypic characteristics of the original tissue such as cell-type diversity. Here, we provide protocols on how to label intestinal (cancer) stem cells by integrating the stem cell ASCL2 reporter (STAR) into human and mouse genomes via two different strategies: (1) lentiviral transduction or (2) transposon-based integration. Organoid technology, in combination with the user-friendly nature of STAR, will facilitate basic research in human and mouse adult stem cell biology. For complete details on the use and execution of this protocol, please refer to Oost et al. (2018).

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Sigma-Aldrich
ダルベッコ改変イーグル培地(高グルコース), With 4500 mg/L glucose, L-glutamine, sodium pyruvate, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
N-アセチル-L-システイン, BioReagent, suitable for cell culture
Sigma-Aldrich
ニコチンアミド, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
ピューロマイシン 二塩酸塩 Streptomyces alboniger由来, ≥98% (HPLC), powder
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臭化ヘキサジメトリン, ≥95%
Sigma-Aldrich
トリプシンインヒビター Glycine max(ダイズ)由来, lyophilized powder