CD244 expression represents functional decline of murine hematopoietic stem cells after in vitro culture

Shuhei Koide; Valgardur Sigurdsson; Visnja Radulovic; Kiyoka Saito; Zhiqian Zheng; Stefan Lang; Shamit Soneji; Atsushi Iwama; Kenichi Miharada

doi:10.1016/j.isci.2021.103603

CD244 expression represents functional decline of murine hematopoietic stem cells after in vitro culture

iScience. 2021 Dec 10;25(1):103603. doi: 10.1016/j.isci.2021.103603. eCollection 2022 Jan 21.

Authors

Shuhei Koide^{1

2}, Valgardur Sigurdsson¹, Visnja Radulovic¹, Kiyoka Saito^{1

3}, Zhiqian Zheng², Stefan Lang⁴, Shamit Soneji⁴, Atsushi Iwama², Kenichi Miharada^{1

3}

Affiliations

¹ Division of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden.
² Division of Stem Cell and Molecular Medicine, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 108-0071 Tokyo, Japan.
³ International Research Center for Medical Sciences, Kumamoto University, 860-0811 Kumamoto, Japan.
⁴ StemTherapy Bioinformatics Core Facility, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden.

Abstract

Isolation of long-term hematopoietic stem cell (HSC) is possible by utilizing flow cytometry with multiple cell surface markers. However, those cell surface phenotypes do not represent functional HSCs after in vitro culture. Here we show that cultured HSCs express mast cell-related genes including Cd244. After in vitro culture, phenotypic HSCs were divided into CD244^- and CD244⁺ subpopulations, and only CD244^- cells that have low mast cell gene expression and maintain HSC-related genes sustain reconstitution potential. The result was same when HSCs were cultured in an efficient expansion medium containing polyvinyl alcohol. Chemically induced endoplasmic reticulum (ER) stress signal increased the CD244⁺ subpopulation, whereas ER stress suppression using a molecular chaperone, TUDCA, decreased CD244⁺ population, which was correlated to improved reconstitution output. These data suggest CD244 is a potent marker to exclude non-functional HSCs after in vitro culture thereby useful to elucidate mechanism of functional decline of HSCs during ex vivo treatment.

Keywords: Biological sciences; Cell biology; Immunology; Stem cells research.