title: Retinoic acid induces mouse bone marrow-derived CD15⁺, Oct4⁺ and CXCR4⁺ stem cells into male germ-like cells in a two-dimensional cell culture system.
abstract: We have examined the effect of retinoic acid (RA) on differentiation of bone marrow-derived CD15(+) , Oct4(+) and CXCR4(+) cells into male germ cells. Bone marrow stem cells (BMSCs) were isolated from the femur of 3-4-week-old male C57BL/6 mice. Magnetic-activated cell sorting (MACS) system was used to sort CD15(+) , Oct4(+) and CXCR4(+) cells. RT-PCR was used to follow the expression of pluripotency markers. Sorted CD15(+) , Oct4(+) and CXCR4(+) cells were cultured in an undifferentiated condition on a feeder layer of mitomycin C-inactivated C2C12. The embryoid-like bodies were differentiated into male germ cells by retinoic acid. To identify the expression of male germ specific markers, differentiated cells were analysed by means of reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence staining. RT-PCR and immunofluorescence show that bone marrow-derived CD15(+) , Oct4(+) and CXCR4(+) cells express pluripotency markers, Oct4, Nanog, Rex-1, SOX-2 and AP. The purified CD15(+) , Oct4(+) and CXCR4(+) formed structures like embryoid bodies when plated over a feeder layer; these bodies were alkaline phosphatase positive. When cells were induced by RA, bone marrow-derived CD15(+) , Oct4(+) and CXCR4(+) were positive for Mvh, Dazl, Piwil2, Dppa3 and Stra8, that known molecular markers of male germ cells. Thus RA can induce differentiation of mouse bone marrow-derived CD15(+) , Oct4(+) and CXCR4(+) cells into male germ cells in vitro. Negative results for the gene expression analysis of female germ cells markers, GDF9 and ZP3, confirmed this conclusion.
authors.affiliation: Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Iraj Ragerdi
authors.initials: IR
authors.lastname: Kashani
authors.forename: Amir Hassan
authors.initials: AH
authors.lastname: Zarnani
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.forename: Mir Abbas
authors.initials: MA
authors.lastname: Abdolvahabi
authors.forename: Karim
authors.initials: K
authors.lastname: Nayernia
authors.forename: Reza
authors.initials: R
authors.lastname: Shirazi
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone; D053487:DEAD-box RNA Helicases / Q000378:metabolism; D004527:Egg Proteins / Q000096:biosynthesis; D005260:Female; D005647:Fucosyltransferases / Q000378:metabolism; D055429:Growth Differentiation Factor 9 / Q000096:biosynthesis; D018398:Homeodomain Proteins / Q000096:biosynthesis; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D010063:Ovum; D016601:RNA-Binding Proteins / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000096:biosynthesis; D012097:Repressor Proteins / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000096:biosynthesis; D013094:Spermatozoa / Q000166:cytology*; D014157:Transcription Factors / Q000096:biosynthesis; D014212:Tretinoin / Q000494:pharmacology*; D000072216:Zona Pellucida Glycoproteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D060565:Argonaute Proteins; C514936:CXCR4 protein, mouse; D002868:Chromosomal Proteins, Non-Histone; C508515:DAZL protein, mouse; C462180:Dppa3 protein, mouse; D004527:Egg Proteins; C525898:Gdf9 protein, mouse; D055429:Growth Differentiation Factor 9; D018398:Homeodomain Proteins; D008562:Membrane Glycoproteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C448521:Piwil2 protein, mouse; C494174:Pou5f1 protein, mouse; D016601:RNA-Binding Proteins; D019718:Receptors, CXCR4; D011956:Receptors, Cell Surface; D012097:Repressor Proteins; C513672:Zfp42 protein, mouse; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; C102019:Stra8 protein, mouse; D014157:Transcription Factors; D000072216:Zona Pellucida Glycoproteins; C000606521:Zp3 protein, mouse; D014212:Tretinoin; D005647:Fucosyltransferases; C541914:Fut4 protein, mouse; C090864:Ddx4 protein, mouse; D053487:DEAD-box RNA Helicases
keywords: bone marrow stem cells; differentiation; male germ cells; retinoic acid