The Effect of Mesenchymal Stem Cell-Derived Microvesicles on Differentiation of Umbilical Cord Blood-Derived Cd34+ Cells Toward Myeloid Lineage Publisher



Shamsasenjan K^{1, 2} ; Timari H² ; Saleh M³
Source: Gene Reports Published:2022

Abstract

Objective: Mesenchymal stem cells (MSCs) are well known for their supportive role in the hematopoiesis and improving hematopoietic engraftment. MSCs exert their effects on the hematopoietic stem cells (HSCs) by cell-to-cell communications, secretion of different growth factors and microvesicles (MVs). MSC-MVs contain different biological substances that some of them such as growth factors, miRNA, and siRNA have regulatory effects in the hematopoiesis. Therefore, in the present study, we have studied the effect of MSC-MVs on the differentiation of umbilical cord blood-derived CD34+ cells (UCB-derived CD34+ cells) toward myeloid lineage. Materials & methods: In this experimental study, UCB-derived CD34+ cells were cultured in the IMDM medium enriched with 10% FBS in the presence of MSC-MVs that isolated from the supernatant of MSCs culture by utilizing differential ultracentrifugation. The culture conditions of CD34+ cells performed in three groups: (1) Culture in the presence of G-CSF and IL-3 cytokines as a control group. (2) Culture in the presence of mentioned cytokines and 10 μg/ml of MVs concentration as an MV1 group. (3) Culture in the presence of mentioned cytokines and 20 μg/ml of MVs concentration as an MV2 group. Finally, after 72 h incubation, CD34+ cells differentiation toward myeloid lineage was investigated by utilizing flow cytometry (CD13 and CD33 markers) and Real Time-PCR (RUNX1, C/EBP-α, PU.1, and G-CSFR genes). Result: The expression of myeloid lineage-specific CD markers (CD13 and CD33) in the presence of the different concentrations of MVs was higher compared to the control group but increased expression of CD13 was not significant. In addition, the expression of myeloid lineage-specific genes (RUNX1, C/EBP-α, PU.1, and G-CSFR) was normalized with the GAPDH gene as the internal control. The expression of all of these genes in the presence of the different concentrations of MVs was higher compared to the control group but increased expression of PU.1 was not significant. Conclusion: This study provided cues of increased differentiation of UCB-derived CD34+ cells toward myeloid lineage in response to MSC-MVs. Therefore, MSC-MVs play a critical role in HSC fate. © 2021