Pluripotent Male Germline Stem Cells and Their Potential in Generation of Transgenic Animals

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2010 32 5 0855-0859 http / /xuebao. jxau. edu. cn Acta Agriculturae Universitatis Jiangxiensis E - mail ndxb7775@ sina. com * 300071 spermatogonial stem cells SSCs Q2 A 1000-2286 2010 05-0855 - 05 Pluripotent Male Germline Stem Cells and Their Potential in Generation of Transgenic Animals LI Zhi-guo LIU Lin * College of Life Sciences Nankai university Tianjin 300071 China 2010-09 - 25 973 2009ZX08006 - O1OB 1985 * 1964 1984 1987 1990 9 1993 7 1994-1996 Babraham 1996 1998 1998 2004 1998 2009 MBL 2003 2007 Nature Nature Cell Biology Nature Biotechnology Proc Natl Acad Sci USA Stem Cells EMBO Reports Human Molecular Genetics Developmental Biology Biology of Reproduction SCI 60 2003-973 ips 973

856 32 Abstract Spermatogonial stem cells SSCs have the ability of self - renewal and differentiation and are the unique adult stem cells in males that can transmit genetic messages to offspring. The establishment and gradual improvement of culture systems for SSCs in mice not only allow a good understanding of regulatory mechanisms underlying proliferation and differentiation of SSCs but also suggest that SSCs have unique advantages in regenerative medicine and generation of transgenic animals. It might be safer to convert SSCs to embryonic stem cells - like pluripotent germ line stem cells compared to virus vector transduced induced pluripotent stem ips cells. Attempts have been made to produce transgenic mice and rats by allogenic transplantation of SSCs. This review will discuss current research progress of pluripotent germline stem cells and challenges and possible solutions of producing large domestic transgenic animals using those stem cells. Key words spermatogonial stem cells pluripotent germ line stem cells transgenic animals 1 PGCs Sertoli seminiferous cords PGCs gonocytes 1 2 2 6 d 3-4 5 spermatocyte 2 ES - like 6-7 2003 Shinohara 8 2004 ES - like 6 2006 Guan 7 ES - like 2008 Conrad 9 LIF LIF + GD- NF 3 ~ 4 STAT3 CD9 KLF4 OTEX VASA POU6F1 DAZL OCT4 SOX2 e - cadherin NANOG GDF3 10-12 13-14

5 857 1 2 1 Spermatogonial stem cells are converted into pluripotent germline stem cells for which genomic DNA can be modified and then used to obtain transgenic offspring by embryo injection and transplantation. 2 Spermatogonial stem cells are cultured in vitro and then genome is modified. These cells are injected into the testis of recipients to generate transgenic offspring by mating the recipients with normal female mice. Fig. 1 1 Ways to generate transgenic animals using spermatogonial stem cells 3 15 16 DNA ips ips 17 ips 3 18-21 2 1 6-7 22

858 32 23-24 SSCs GDNF Ras /ERK1 /2 25 BMP4 cdh I kit 26 ERK1 /2 BMP4 4 ips ips 1 Dym M. Expression of c - kit receptor and its autophosphorylation in immature rat type A spermatogonia J. Biol Reprod 1995 52 1 8-19. 2 Paniagua R Nistal M. Morphological and histometric study of human spermatogonia from birth to the onset of puberty J. J Anat 1984 139 Pt 3 535-552. 3 Wu X. Prepubertal human spermatogonia and mouse gonocytes share conserved gene expression of germline stem cell regulatory molecules J. Proc Natl Acad Sci U S A 2009 106 51 21672-21677. 4 Culty M. Gonocytes the forgotten cells of the germ cell lineage J. Birth Defects Res C Embryo Today 2009 87 1 1-26. 5 Wu Z. Capacity for stochastic self - renewal and differentiation in mammalian spermatogonial stem cells J. J Cell Biol 2009 187 4 513-524. 6 Kanatsu - Shinohara M. Generation of pluripotent stem cells from neonatal mouse testis J. Cell 2004 119 7 1001-1012.

5 859 7 Guan K. Pluripotency of spermatogonial stem cells from adult mouse testis J. Nature 2006 440 7088 1199-1203. 8 Kanatsu - Shinohara M. Long - term proliferation in culture and germline transmission of mouse male germline stem cells J. Biol Reprod 2003 69 2 612-616. 9 Conrad S. Generation of pluripotent stem cells from adult human testis J. Nature 2008 456 7220 344-349. 10 Kuijk E W Colenbrander B Roelen B A. The effects of growth factors on in vitro - cultured porcine testicular cells J. Reproduction 2009 138 4 721-731. 11 Aponte P M. Propagation of bovine spermatogonial stem cells in vitro J. Reproduction 2008 136 5 543-557. 12 Cheng G S Feng T. Studies on spermatogonial stem cells cultured in vitro of Wuzhishan Mini Porcine J. 2006 22 4 689-93. 13 Ryu B Y. Conservation of spermatogonial stem cell self - renewal signaling between mouse and rat J. Proc Natl Acad Sci U S A 2005 102 40 14302-14307. 14 Kanatsu - Shinohara M. Long - term culture of male germline stem cells from hamster testes J. Biol Reprod 2008 78 4 611-617. 15 Robertson E B A Kuehn M Evans M. Germ - line transmission of genes introduced into cultured pluripotential cells by retroviral vector J. Nature 1986 323 6087 445-458. 16 Kawamata M Ochiya T. Generation of genetically modified rats from embryonic stem cells J. Proc Natl Acad Sci U S A 2010 107 32 14223-14228. 17 Kou Z. Mice cloned from induced pluripotent stem cells ipscs J. Biol Reprod 2010 83 2 238-243. 18 Nagano M. Retrovirus - mediated gene delivery into male germ line stem cells J. FEBS Lett 2000 475 1 7-10. 19 Nagano M. Transgenic mice produced by retroviral transduction of male germ - line stem cells J. Proc Natl Acad Sci U S A 2001 98 23 13090-13095. 20 Kanatsu - Shinohara M Toyokuni S Shinohara T. Transgenic mice produced by retroviral transduction of male germ line stem cells in vivo J. Biol Reprod 2004 71 4 1202-1207. 21 Hamra F K. Production of transgenic rats by lentiviral transduction of male germ - line stem cells J. Proc Natl Acad Sci U S A 2002 99 23 14931-14936. 22 Ko K A. - B M Kim J Stehling M et al. Conversion of adult mouse unipotent germline stem cells into pluripotent stem cells J. Nat Protoc 2010 5 5 921-928. 23 Brinster R L Zimmermann J W Spermatogenesis following male germ - cell transplantation J. Proc Natl Acad Sci U S A 1994 91 24 11298-11302. 24 Brinster R L Avarbock M R Germline transmission of donor haplotype following spermatogonial transplantation J. Proc Natl Acad Sci U S A 1994 91 24 11303-11307. 25 He Z. Gdnf upregulates c - Fos transcription via the Ras /Erk1 /2 pathway to promote mouse spermatogonial stem cell proliferation R. Stem Cells 2008 26 1 266-278. 26 Carlomagno G. BMP4 - Induced Differentiation of a Rat Spermatogonial Stem Cell Line Causes Changes in Its Cell Adhesion Properties R. Biol Reprod 2010. on line.