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      Isolation and Characterization of Pluripotent Human Spermatogonial Stem Cell-Derived Cells

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          Abstract

          Several reports have documented the derivation of pluripotent cells (multipotent germline stem cells) from spermatogonial stem cells obtained from the adult mouse testis. These spermatogonia-derived stem cells express embryonic stem cell markers and differentiate to the three primary germ layers, as well as the germline. Data indicate that derivation may involve reprogramming of endogenous spermatogonia in culture. Here, we report the derivation of human multipotent germline stem cells (hMGSCs) from a testis biopsy. The cells express distinct markers of pluripotency, form embryoid bodies that contain derivatives of all three germ layers, maintain a normal XY karyotype, are hypomethylated at the H19 locus, and express high levels of telomerase. Teratoma assays indicate the presence of human cells 8 weeks post-transplantation but limited teratoma formation. Thus, these data suggest the potential to derive pluripotent cells from human testis biopsies but indicate a need for novel strategies to optimize hMGSC culture conditions and reprogramming.

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          Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells.

          G Martin (1981)
          This report describes the establishment directly from normal preimplantation mouse embryos of a cell line that forms teratocarcinomas when injected into mice. The pluripotency of these embryonic stem cells was demonstrated conclusively by the observation that subclonal cultures, derived from isolated single cells, can differentiate into a wide variety of cell types. Such embryonic stem cells were isolated from inner cell masses of late blastocysts cultured in medium conditioned by an established teratocarcinoma stem cell line. This suggests that such conditioned medium might contain a growth factor that stimulates the proliferation or inhibits the differentiation of normal pluripotent embryonic cells, or both. This method of obtaining embryonic stem cells makes feasible the isolation of pluripotent cells lines from various types of noninbred embryo, including those carrying mutant genes. The availability of such cell lines should made possible new approaches to the study of early mammalian development.
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            Reprogramming of human somatic cells to pluripotency with defined factors.

            In-Hyun Park, Rui Zhao, Jason A. West … (2008)
            Pluripotency pertains to the cells of early embryos that can generate all of the tissues in the organism. Embryonic stem cells are embryo-derived cell lines that retain pluripotency and represent invaluable tools for research into the mechanisms of tissue formation. Recently, murine fibroblasts have been reprogrammed directly to pluripotency by ectopic expression of four transcription factors (Oct4, Sox2, Klf4 and Myc) to yield induced pluripotent stem (iPS) cells. Using these same factors, we have derived iPS cells from fetal, neonatal and adult human primary cells, including dermal fibroblasts isolated from a skin biopsy of a healthy research subject. Human iPS cells resemble embryonic stem cells in morphology and gene expression and in the capacity to form teratomas in immune-deficient mice. These data demonstrate that defined factors can reprogramme human cells to pluripotency, and establish a method whereby patient-specific cells might be established in culture.
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              Retinoic acid regulates sex-specific timing of meiotic initiation in mice.

              Jana Koubova, Douglas B Menke, Qing Zhou … (2006)
              In mammals, meiosis is initiated at different time points in males and females, but the mechanism underlying this difference is unknown. Female germ cells begin meiosis during embryogenesis. In males, embryonic germ cells undergo G0/G1 mitotic cell cycle arrest, and meiosis begins after birth. In mice, the Stimulated by Retinoic Acid Gene 8 (Stra8) has been found to be required for the transition into meiosis in both female and male germ cells. Stra8 is expressed in embryonic ovaries just before meiotic initiation, whereas its expression in testes is first detected after birth. Here we examine the mechanism underlying the sex-specific timing of Stra8 expression and meiotic initiation in mice. Our work shows that signaling by retinoic acid (RA), an active derivative of vitamin A, is required for Stra8 expression and thereby meiotic initiation in embryonic ovaries. We also discovered that RA is sufficient to induce Stra8 expression in embryonic testes and in vitamin A-deficient adult testes in vivo. Finally, our results show that cytochrome p450 (CYP)-mediated RA metabolism prevents premature Stra8 expression in embryonic testes. Treatment with an inhibitor specific to RA-metabolizing enzymes indicates that a cytochrome p450 from the 26 family (CYP26) is responsible for delaying Stra8 expression in embryonic testes. Sex-specific regulation of RA signaling thus plays an essential role in meiotic initiation in embryonic ovaries and precludes its occurrence in embryonic testes. Because RA signaling regulates Stra8 expression in both embryonic ovaries and adult testes, this portion of the meiotic initiation pathway may be identical in both sexes.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Stem Cells
                Journal ID (publisher-id): stem
                Title: Stem Cells (Dayton, Ohio)
                Publisher: Wiley Subscription Services, Inc., A Wiley Company
                ISSN (Print): 1066-5099
                ISSN (Electronic): 1549-4918
                Publication date (Print): January 2009
                Volume: 27
                Issue: 1
                Pages: 138-149
                Affiliations
                [a ]simpleInstitute for Stem Cell Biology and Regenerative Medicine, Department of Obstetrics and Gynecology, Stanford University School of Medicine Palo Alto, California, USA
                [b ]simpleCenter of Reproductive Medicine and Andrology, University of Muenster Muenster, Germany
                [c ]simpleCenter for Reproductive Sciences, University of California San Francisco, San Francisco, California, USA
                [d ]simpleDepartment of Urology, University of California San Francisco, San Francisco, California, USA
                [e ]simpleSheba Medical Center, Tel Hashomer Israel
                Author notes
                Correspondence: Paul J. Turek, M.D., University of California, San Francisco, San Francisco, California 94043, USA. Telephone: 415-392-3200; e-mail: pturek@ 123456urology.ucsf.edu
                Renee A. Reijo-Pera, Ph.D., Stanford University School of Medicine, Palo Alto, California 94304-5542, USA. Telephone: 650-725-3803; Fax: 650-736-2961

                Author contributions: N.K.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; J.M. and S.S.: collection and/or assembly of data, data analysis and interpretation; H.N.N., S.C., and C.N.: collection and/or assembly of data, data analysis and interpretation, manuscript writing; J.G.: manuscript writing; P.J.T.: conception and design, provision of study material or patients, manuscript writing; R.A.R.-P.: conception and design, financial support, administrative support, data analysis and interpretation, manuscript writing, final approval of manuscript.

                Disclosure of potential conflicts of interest is found at the end of this article.

                First published online in S tem C ells Express October 16, 2008; available online without subscription through the open access option.

                Article
                DOI: 10.1634/stemcells.2008-0439
                PMC ID: 2729695
                PubMed ID: 18927477
                SO-VID: 18e3242a-7f35-4b58-b270-3dd222bbed70
                Copyright © Copyright © 2009 AlphaMed Press
                License:

                Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.

                History
                Date received : 05 May 2008
                Date accepted : 22 September 2008
                Categories
                Subject: Tissue-Specific Stem Cells

                ScienceOpen disciplines: Molecular medicine
                Keywords: adult stem cells,human embryonic stem cells,germline stem cells,spermatogonia,testis biopsy
                Data availability:
                ScienceOpen disciplines: Molecular medicine
                Keywords: adult stem cells, human embryonic stem cells, germline stem cells, spermatogonia, testis biopsy

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