Isolation of human fetal intestinal cells for single-cell RNA sequencing

STAR Protoc. 2021 Oct 22;2(4):100890. doi: 10.1016/j.xpro.2021.100890. eCollection 2021 Dec 17.

Abstract

The intestine has a large number of cell types. Thus, digestion of pure and viable populations is necessary for downstream techniques including single-cell RNA sequencing. We outline a protocol to isolate both epithelial and non-epithelial cells from human fetal samples at high viability, which was used to produce a full thickness atlas of intestinal cells across human development. This protocol can also be adapted to adult endoscopy and surgical specimens. For details on the use of this protocol, please refer to Fawkner-Corbett et al. (2021).

Keywords: Cell Biology; Cell isolation; Developmental biology; Flow Cytometry/Mass Cytometry; Single Cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Fetus / cytology*
  • Fetus / metabolism
  • Humans
  • Intestines / cytology*
  • Intestines / metabolism
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Analysis, RNA / methods*
  • Single-Cell Analysis / methods*

Substances

  • RNA, Messenger