Processing Human Thymic Tissue for Single Cell RNA-Seq

Justin Le; Vi Luan Ha; Annie Luong; Chintan Parekh

doi:10.1016/j.xpro.2020.100090

Processing Human Thymic Tissue for Single Cell RNA-Seq

STAR Protoc. 2020 Aug 24;1(2):100090. doi: 10.1016/j.xpro.2020.100090. eCollection 2020 Sep 18.

Authors

Justin Le¹, Vi Luan Ha^{1

2}, Annie Luong¹, Chintan Parekh¹

Affiliations

¹ Cancer and Blood Disease Institute, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA.
² Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, CA 90027, USA.

Abstract

Single cell RNA sequencing of human thymic cells is dependent on isolation of highly pure and viable cell populations. This protocol describes the isolation of CD34⁺ progenitor and more differentiated CD34^- fractions from post-natal thymic tissue to study thymopoiesis. CD34⁺ cells represent <1% of thymic cells, so this protocol uses magnetic- followed by fluorescence-activated cell separation to isolate highly enriched CD34⁺ cells. For complete details on the use and execution of this protocol, please refer to Le et al. (2020).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antigens, CD34 / immunology
Antigens, CD34 / isolation & purification
Bone Marrow Cells
Cell Differentiation
Cell Separation / methods*
Cells, Cultured
Humans
RNA / genetics
RNA / isolation & purification
RNA-Seq / methods*
Single-Cell Analysis / methods*
Thymus Gland / metabolism
Thymus Gland / physiology

Substances

Antigens, CD34
RNA