title: Differentiation of bone marrow-derived stage-specific embryonic antigen 1 positive pluripotent stem cells into male germ cells.
abstract: Studies published in recent years have changed the outlook on sterility and germ cell development by producing gametes from stem cells. In present study, a novel approach on differentiation of bone marrow-derived stage-specific embryonic antigen 1 positive (SSEA-1+) pluripotent stem cells into male germ cells has been addressed. SSEA-1+stem cells were separated from murine bone marrow using magnetic-activated cell sorting (MACS) system and propagated on a feeder layer cells. To evaluate the pluripotency characteristic of the purified cells, they were differentiated toward cells of three germ layers. Later the SSEA-1+stem cells were induced to differentiate along male germ cell lineage with retinoic acid. Flowcytometric analysis of SSEA-1+stem cells revealed purity of about 62% which increased to 91% after cultivation over feeder cells. Expression of specific transcripts of Oct4, SSEA-1, Nanog, Dppa3, fragilis, Rex-1, SOX-2, and alkaline-phosphatase and immunofluorescence evaluation of Oct4 and SSEA-1 expression showed the differentiation of purified stem cells toward the cells of three germ layers. Differentiation potential of purified cells was positively evidenced by expression markers specific for primordial germ cells, spermatogonial stem cells and spermatogonia including Mvh, fragilis, Dppa3, Stra8, DAZL, Piwil2, β1, and α6-integrins as well as meiotic-specific marker SYCP3. Our results showed that SSEA-1+pluripotent stem cells are able to differentiate into male germ cells. The results of the present study are encouraging enough to merit further investigation, provide a new hope for those suffering from infertility and introduce a novel platform for research on germ cell development.
authors.affiliation: Cellular and Molecular Research Center Iran University of Medical Sciences, Tehran, Iran.|Department of Anatomical Sciences School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Shirazi
authors.affiliation: Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.|Immunology Research Center Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Amir Hassan
authors.initials: AH
authors.lastname: Zarnani
authors.affiliation: Faculty of Medical Sciences, Department of Hematology, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: GENEOCELL, Institute of advanced bimolecular and cellular technologies, Montreal, Canada.
authors.forename: Karim
authors.initials: K
authors.lastname: Nayernia
authors.affiliation: Department of Anatomical Sciences School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Iraj
authors.initials: I
authors.lastname: Ragerdi Kashani
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002478:Cells, Cultured; D016256:Lewis X Antigen / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010006:Osteoblasts / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology*; D012583:Schwann Cells / Q000166:cytology; D013091:Spermatogenesis / Q000502:physiology*; D013093:Spermatogonia / Q000166:cytology*; D013094:Spermatozoa / Q000166:cytology* / Q000254:growth & development; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D016256:Lewis X Antigen; D014212:Tretinoin
keywords: fertility; gametogenesis; retinoic acid; spermatogonial stem cells