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

February 1 2022

Lab: Kenichi Miharada

Paper information

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

Shusei Koide, Valgardur Sigurdsson, Visnja Radulovic, Kiyoka Saito, Zhiqian Zheng, Stefan Lang, Shamit Soneji,

Atsushi Iwama and Kenichi Miharada

iScience January 21, 2022

doi: https://doi.org/10.1016/j.isci.2021.103603

Highlishts

• Murine HSCs up-regulate mast cell-related genes including Cd244 during in vitro culture
• Long-term HSCs after in vitro culture are enriched in CD244⁻CD48⁻KSL population
• Induction of unfolded protein response is involved in the increase of CD244⁺HSC

Summary

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.