Effect of Recombinant Human Erythropoietin On the Stemness of Bone Marrow-derived Mesenchymal Stem Cells in vitro

Abstract

The purpose of this study was to investigate the effects of the recombinant human erythropoietin (rhEPO) on proliferative and multi-differentiation potential of the bone marrow-derived mesenchymal stem cells (MSCs). The MSCs were isolated primarily from bone marrow of adult rat and purified at increasing passage. A purified population of MSCs can be obtained about 2 weeks after the initiation of culture. After three passages (P3-MSCs), bone marrow-derived adherent cells were identified, then different concentrations of rhEPO (0.1, 1, 5, 10, 100 U/ml) was added into the Passage-3 cells which had been identified. The expression of the surface markers in adherent cells was detected by the flow cytometry. The mRNA levels of transcription factors OCT4, SOX2, Nanog and TERT were measured by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that CD29 and CD90 were positive in MSCs, but not CD33, CD44 and CD45, and the cells could differentiate into multiple lineages such as osteocytes and adipocytes. The expression of OCT4, SOX2, TERT, Nanog mRNA were up-regulated by the treatment of EPO. The effect of EPO was the most obvious when its concentration was 5U/mL after 12h. we conclude that MSCs can not only perserve characteristics of stem cells but also maintain its multi-lineage differentiation potential after appropriate treatment of EPO.

Pleiotropic effects of Erythropoietin. Influence of Erythropoietin on processes of mesenchymal stem cells differentiation

“…Endogenous erythropoietin and recombinant erythropoietin are identical, except for some minor differences in glycosylation (Koury andBondurant 1992, Rölfing 2014). Human recombinant erythropoietin was first characterised as a hematopoietic factor (Ye et al 2010). It was shown that this substance stimulated differentiation and proliferation of erythroid precursor cells (Broudy et al 1991, Ye et al 2010.…”

“…Human recombinant erythropoietin was first characterised as a hematopoietic factor (Ye et al 2010). It was shown that this substance stimulated differentiation and proliferation of erythroid precursor cells (Broudy et al 1991, Ye et al 2010. Recombinant forms of erythropoietin are epoetin-alfa (human erythropoietin which is produced in cell culture using recombinant DNA technology), epoetin-beta (which is a synthetic recombinant form of EPO) and long-term acting analogue darbepoetin-alfa (which is also a synthetic form of erythropoietin) (Arcasoy 2008, Uversky andRedwan 2017).…”

“…There were identified some properties of recombinant erythropoietin in regulation of genes expression and their function similar to vascular endothelial growth factor (VEGF), main angiogenic factor (Suk et al 2008). It is known that erythropoietin is able to stimulate proliferation of endothelial cells in vitro and mobilize endothelial progenitor cells from bone marrow (Bahlmann et al 2003, Bahlmann et al 2004, Heeschen et al 2003, Klopsch et al 2009, increase adhesive and proliferative characteristics of circulating endothelial progenitor cells (George et al 2005, Klopsch et al 2009 and induce neovascularization of ischemic tissue in vivo (Ye et al 2010). Under the influence of erythropoietin, embryonic stem cells can differentiate into cardiomyocytes; erythropoietin also facilitates proliferation of cardiomyocytes (Broudy et al 1991, Sachinidis et al 2002, Ye et al 2010.…”

“…It is known that erythropoietin is able to stimulate proliferation of endothelial cells in vitro and mobilize endothelial progenitor cells from bone marrow (Bahlmann et al 2003, Bahlmann et al 2004, Heeschen et al 2003, Klopsch et al 2009, increase adhesive and proliferative characteristics of circulating endothelial progenitor cells (George et al 2005, Klopsch et al 2009 and induce neovascularization of ischemic tissue in vivo (Ye et al 2010). Under the influence of erythropoietin, embryonic stem cells can differentiate into cardiomyocytes; erythropoietin also facilitates proliferation of cardiomyocytes (Broudy et al 1991, Sachinidis et al 2002, Ye et al 2010. Erythropoietin is able to influence differentiation of neuronal stem cells into neuronal cells (Chen et al 2009, Kretz et al 2005, Ye et al 2010.…”

An overview on different strategies for the stemness maintenance of MSCs

“…The recombinant human erythropoietin (rhEPO) can not only keep features of stem cells but also preserve its multi-lineage differentiation ability after suitable EPO treatment. But extra investigation on the involved mechanism can lead to a better understanding between EPO and MSC stemness maintenance (Ye et al 2010).…”