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
Affiliations
Shuhei Koide
Division of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden; 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
Valgardur Sigurdsson
Division of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden
Visnja Radulovic
Division of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden
Kiyoka Saito
Division of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden; International Research Center for Medical Sciences, Kumamoto University, 860-0811 Kumamoto, Japan
Zhiqian Zheng
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
Stefan Lang
StemTherapy Bioinformatics Core Facility, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden
Shamit Soneji
StemTherapy Bioinformatics Core Facility, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden
Atsushi Iwama
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
Kenichi Miharada
Division of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden; International Research Center for Medical Sciences, Kumamoto University, 860-0811 Kumamoto, Japan; Corresponding author
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.
