Fluidigm Based Single-cell Gene Expression Library Preparation from Patient-derived Small Intestinal Organoids

Kohei Suzuki; Ryuichi Okamoto

doi:10.21769/BioProtoc.3775

Fluidigm Based Single-cell Gene Expression Library Preparation from Patient-derived Small Intestinal Organoids

Bio Protoc. 2020 Oct 5;10(19):e3775. doi: 10.21769/BioProtoc.3775.

Authors

Kohei Suzuki¹, Ryuichi Okamoto^{2

3}

Affiliations

¹ Department of Internal Medicine, Division of Digestive and Liver Diseases, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
² Department of Gastroenterology and Hepatology.
³ Center for Stem Cell and Regenerative Medicine, Graduate School, Tokyo Medical and Dental University, Tokyo, 113-8519, Japan.

Abstract

In this protocol, we describe our methods to isolate crypts from patients' biopsy samples and to culture human intestinal stem cells as it's called "organoid." Beyond that, we describe how to dissociate organoids cells into single cells for single-cell analysis as a further application. This protocol should provide investigators sufficient tools to generate human organoids from biopsy samples and to accomplish a stable in-vitro assay system.

Keywords: Endoscopic biopsy; Intestinal stem cell; Multiplex PCR; Organoid; Single-cell analysis.