Molecular signature of erythroblast enucleation in human embryonic stem cells

Abstract

While enucleation is a critical step in the terminal differentiation of human red blood cells, the molecular mechanisms underlying this unique process remain unclear. To investigate erythroblast enucleation, we studied the erythroid differentiation of human embryonic stem cells (hESCs), which provide a unique model for deeper understanding of the development and differentiation of multiple cell types. First, using a two-step protocol, we demonstrated that terminal erythroid differentiation from hESCs is directly dependent on the age of the embryoid bodies. Second, by choosing hESCs in two extreme conditions of erythroid culture, we obtained an original differentiation model which allows one to study the mechanisms underlying the enucleation of erythroid cells by analyzing the gene and miRNA (miR) expression profiles of cells from these two culture conditions. Third, using an integrated analysis of mRNA and miR expression profiles, we identified five miRs potentially involved in erythroblast enucleation. Finally, by selective knockdown of these five miRs we found miR-30a to be a regulator of erythroblast enucleation in hESCs.

High-Efficiency Serum-Free Feeder-Free Erythroid Differentiation of Human Pluripotent Stem Cells Using Small Molecules

“…The recent report of Rouzbeh et al [41] demonstrated that it might be possible to increase the percentage of cells undergoing enucleation by reducing the expression of micro RNA30a (MIR30a); however, there was very little, if any, cell expansion in their system. Furthermore, knockdown of an MIR is a difficult strategy to use routinely, whereas identification of the MIR30a targets and development of small molecules effectors would increase the utility of the method and could be easily tested in the high-yield, highly defined differentiation system reported here.…”

Activation of KLF1 Enhances the Differentiation and Maturation of Red Blood Cells from Human Pluripotent Stem Cells

“…Scalable, clinically compatible protocols to produce erythroid cells from hPSCs have been developed but progress to translation has been hampered by the lack of terminal maturation of the resultant cell. In contrast to RBCs generated in vitro from adult bone marrow or mobilised peripheral blood CD34 1 progenitors cells, erythroid cells produced from both hESCs and iPSCs have a fragile morphology, a poor enucleation rates and express embryonic and foetal rather than adult globin [1][2][3][4][5][6][7].…”

MiR‐30a increases MDSC differentiation and immunosuppressive function by targeting SOCS3 in mice with B‐cell lymphoma

“…MDSC expansion, function and migration are controlled by miR-494 [19], miR-155 [17,20,21], miR-21 [17], miR-210 [22] and miR-9 [16]. Recent studies have reported that miR-30 can regulate the differentiation and function of many cell types, such as mesenchymal stem cells [23,24] and human embryonic stem cells [25]. Our previous study also found that miR-30a attenuated the immunosuppressive functions of IL-1b-elicited mesenchymal stem cells [26].…”