Current Issue

Volume 20, Number 2, Summer 2018, Serial Number: 78 Pages: 195-203

MicroRNA Microarray Profiling during Megakaryocyte Differentiation of Cord Blood CD133+ Hematopoietic Stem Cells


Mohammad Houshmand, M.Sc, 1, 2, Mozhde Nakhlestani Hagh, M.D, 1, Masoud Soleimani, Ph.D, 3, Amir Ali Hamidieh, M.D., 4, Saeed Abroun, Ph.D., 3, Mahin Nikougoftar Zarif, Ph.D., 1, 5, *,
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
Department of Clinical and Biological Sciences, University of Turin, San Luigi Gonzaga Hospital, Orbassano, Italy
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Children’s Medical Center, Tehran University of Medical Science, Tehran, Iran
HSCT Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
*Corresponding Address: P.O.Box: 146651157 Blood Transfusion Research Center High Institute for Research and Education in Transfusion Medicine Tehran Iran Email:nikougoftar@ibto.ir

Abstract

Objective

In order to clarify the role of microRNAs (miRNA) in megakaryocyte differentiation, we ran a microRNA microarray experiment to measure the expression level of 961 human miRNA in megakaryocytes differentiated from human umbilical cord blood CD133+ cells.

Materials and Methods

In this experimental study, human CD133+ hematopoietic stem cells were collected from three human umbilical cord blood (UCB) samples, and then differentiated to the megakaryocytic lineage and characterized by flow cytometry, CFU-assay and ploidy analysis. Subsequently, microarray analysis was undertaken followed by quantitative polymerase chain reaction (qPCR) to validate differentially expressed miRNA identified in the microarray analysis.

Results

A total of 10 and 14 miRNAs were upregulated (e.g. miR-1246 and miR-148-a) and down-regulated (e.g. miR- 551b and miR-10a) respectively during megakaryocyte differentiation, all of which were confirmed by qPCR. Analysis of targets of these miRNA showed that the majority of targets are transcription factors involved in megakaryopoiesis.

Conclusion

We conclude that miRNA play an important role in megakaryocyte differentiation and may be used as targets to change the rate of differentiation and further our understanding of the biology of megakaryocyte commitment.