title: Megakaryocytic differentiation of CD133+ hematopoietic stem cells by down-regulation of microRNA-10a.
abstract: MicroRNAs (miRNA), as a class of non-coding RNAs, play a crucial role in normal hematopoiesis. Recent studies on miRNA profiling during hematopoiesis showed miR-10a down-regulation in megakaryocytic differentiation. Here we investigated whether decrease in miR-10a can differentiate umbilical cord blood CD133+ cells to megakaryocytic series. We ectopically induced miR-10a down-regulation by locked nucleic acid anti-miR-10a transfection of CD133+ cells. The megakaryocytic differentiation was evaluated by CD42/CD61 and CD41 surface expression and colonogenic capacity in Megacult media. In addition, real-time polymerase chain reaction was done for detection of miR-10a and its target HoxA1 mRNA. HOXA1 protein expression was detected by flow cytometry as mean fluorescent intensity that shows the antibody reaction proteins. Our findings showed megakaryocytic differentiation of about 28% of umbilical cord blood CD133+ stem cells with bright expression of CD42/CD61 and CD41 in parallel with increase in HoxA1 mRNA and protein level. Colony forming of CD61+ cells in Megacult demonstrated the colonogenic capacity of differentiated progenitor cells. In conclusion, MiR-10a has a role in megakaryocyte differentiation of stem cells via HOXA1 transcription factor targeting.
authors: Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
authors: M Nikougoftar
authors: MN
authors: Zarif
authors: M
authors: M
authors: Soleimani
authors: H
authors: H
authors: Abolghasemi
authors: N
authors: N
authors: Amirizade
authors: E
authors: E
authors: Arefian
authors: A
authors: A
authors: Rahimian
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D015536:Down-Regulation; D005312:Fetal Blood / Q000166:cytology; D005434:Flow Cytometry; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006023:Glycoproteins / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D039661:Integrin beta3 / Q000378:metabolism; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D010455:Peptides / Q000378:metabolism; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000099261:Homeobox A1 Protein
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; D018398:Homeodomain Proteins; D039661:Integrin beta3; D035683:MicroRNAs; D010455:Peptides; D019038:Platelet Glycoprotein GPIb-IX Complex; D040201:Platelet Membrane Glycoprotein IIb; D012333:RNA, Messenger; D014157:Transcription Factors; D000099261:Homeobox A1 Protein; C547183:MIRN10 microRNA, human; C000606023:PROM1 protein, human