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

Abstract

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.

Maternal sterol 27-hydroxylase is crucial for securing fetal development

“…Gene ontology (GO) analysis using ToppFun also revealed that the genes upregulated in KO mat fetal lung cells were mainly implicated in ribosome biogenesis and the translocation of proteins to the ER (Figure 3E). BAs function as a chemical chaperone that inhibits the induction of ER stress, both in vitro and in vivo (Özcan et al, 2011; Miharada et al, 2014; Sigurdsson et al, 2016; Sigurdsson et al, 2020; Koide et al, 2022). However, we failed to observe any induction of ER stress markers (Supplementary Figure S5B), suggesting the involvement of a different mechanism.…”

A culture platform to study quiescent hematopoietic stem cells following genome editing

“…Koide et al. 74 reported that the surface marker phenotype of HSCs (CD48 − LSK cells) cultured under the PVA-based condition reflects their in vivo repopulation capacity. 74 This suggests that retaining the HSC phenotype after genome editing is an alternative approach to maintaining functional HSCs.…”

“… 74 reported that the surface marker phenotype of HSCs (CD48 − LSK cells) cultured under the PVA-based condition reflects their in vivo repopulation capacity. 74 This suggests that retaining the HSC phenotype after genome editing is an alternative approach to maintaining functional HSCs. Notably, re-entry into quiescence requires adjustment of the concentrations of only two essential cytokines for HSCs (SCF and TPO).…”

Hematopoietic stem cells and extramedullary hematopoiesis in the lungs

“…The significance and function of CD150 and CD48 are outlined above. CD244 is a marker of MPPs in humans and mice and is notably absent on healthy HSCs, as its expression marks the functional decline of HSCs in a murine model [43]. Differential expression of these three markers allows for identification of proliferating hematopoietic cells in various developmental stages, including HSCs, MPPs, and restricted progenitors [41].…”