Name: germ cell transplantation and testis tissue xenografting in mice
Uploaded: 2012-02-06T12:00:00
Description: Watch this Scientific Journal Video about Germ Cell Transplantation and Testis Tissue Xenografting in Mice at JoVE.com

Arbete från författarna laboratoriet har fått stöd av USDA / CSREES / NRICGP (2007-35203-18213) NIH / NCRR (2 R01 RR17359-06), NIH / NIEHS (1 R21 ES014856-01A2) och Alberta innovativt - Health Solutions.

DEL A. könsceller transplantation hos möss 1. Beredning av mottagande möss <ol><li> Mottagarna ska vara immunologiskt toleranta (antingen genetiskt anpassade till givare eller immunbrist) till cellerna donator testiklar. </li><li> Mottagarna ska antingen vara naturligt saknar spermatogenesen (t.ex. W / W v möss) eller utarmat av endogena könsceller. Hos könsceller utarmning kan uppnås genom att bestrålning eller kemoterapeutiska droger såsom Busulfan. I detta pro...

<ol>
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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+factors+essential+for+self-renewal+and+expansion+of+mouse+spermatogonial+stem+cells.">Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells.</a> Proceedings of the National Academy of Sciences of the United States of America. 101 (47), 16489-16489 (2004).</li><li>Oatley, J. M., Avarbock, M. R., Telaranta, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identifying+genes+important+for+spermatogonial+stem+cell+self-renewal+and+survival.">Identifying genes important for spermatogonial stem cell self-renewal and survival.</a> Proceedings of the National Academy of Sciences of the United States of America. 103 (25), 9524-9524 (2006).</li><li>Dobrinski, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Germ+cell+transplantation+and+testis+tissue+xenografting+in+domestic+animals.">Germ cell transplantation and testis tissue xenografting in domestic animals.</a> Animal Reproduction Science. 89 (1-4), 137-137 (2005).</li><li>Dobrinski, I., Avarbock, M. R., Brinster, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+germ+cells+from+rabbits+and+dogs+into+mouse+testes.">Transplantation of germ cells from rabbits and dogs into mouse testes.</a> Biology of Reproduction. 61 (5), 1331-1331 (1999).</li><li>Honaramooz, A., Snedaker, A., Boiani, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sperm+from+neonatal+mammalian+testes+grafted+in+mice.">Sperm from neonatal mammalian testes grafted in mice.</a> Nature. 418 (6899), 778-778 (2002).</li><li>Rodriguez-Sosa, J. R., Dobrinski, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+developments+in+testis+tissue+xenografting.">Recent developments in testis tissue xenografting.</a> Reproduction. 138 (2), 187-187 (2009).</li><li>Dobrinski, I., Rathi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ectopic+grafting+of+mammalian+testis+tissue+into+mouse+hosts.">Ectopic grafting of mammalian testis tissue into mouse hosts.</a> Methods in Molecular Biology. 139, 450-450 (2008).</li><li>Rodriguez-Sosa, J. R., Schlatt, S., Dobrinski, I., Seli, E., Agarwal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testicular+tissue+transplantation+for+fertility+preservation.">Testicular tissue transplantation for fertility preservation.</a> Fertility Preservation: Emerging Technologies and Clinical Applications. , 331-331 (2011).</li><li>Rathi, R., Honaramooz, A., Zeng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Germ+cell+fate+and+seminiferous+tubule+development+in+bovine+testis+xenografts.">Germ cell fate and seminiferous tubule development in bovine testis xenografts.</a> Reproduction. 130 (6), 923-923 (2005).</li><li>Rathi, R., Honaramooz, A., Zeng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Germ+cell+development+in+equine+testis+tissue+xenografted+into+mice.">Germ cell development in equine testis tissue xenografted into mice.</a> Reproduction. 131 (6), 1091-1091 (2006).</li><li>Kim, Y., Selvaraj, V., Pukazhenthi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+donor+age+on+success+of+spermatogenesis+in+feline+testis+xenografts.">Effect of donor age on success of spermatogenesis in feline testis xenografts.</a> Reproduction, Fertility, and Development. 19 (7), 869-869 (2007).</li><li>Arregui, L., Rathi, R., Zeng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xenografting+of+adult+mammalian+testis+tissue.">Xenografting of adult mammalian testis tissue.</a> Animal Reproduction Science. 106 (1-2), 65-65 (2008).</li><li>Schlatt, S., Honaramooz, A., Ehmcke, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limited+survival+of+adult+human+testicular+tissue+as+ectopic+xenograft.">Limited survival of adult human testicular tissue as ectopic xenograft.</a> Human Reproduction. 21 (2), 384-384 (2006).</li><li>Shinohara, T., Avarbock, M. R., Brinster, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=beta1-+and+alpha6-integrin+are+surface+markers+on+mouse+spermatogonial+stem+cells.">beta1- and alpha6-integrin are surface markers on mouse spermatogonial stem cells.</a> Proceedings of the National Academy of Sciences of the United States of America. 96 (10), 5504-5504 (1999).</li><li>Kanatsu-Shinohara, M., Toyokuni, S., Shinohara, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD9+is+a+surface+marker+on+mouse+and+rat+male+germline+stem+cells.">CD9 is a surface marker on mouse and rat male germline stem cells.</a> Biology of Reproduction. 70 (1), 70-70 (2004).</li><li>Ryu, B. Y., Orwig, K. E., Oatley, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+aging+and+niche+microenvironment+on+spermatogonial+stem+cell+self-renewal.">Effects of aging and niche microenvironment on spermatogonial stem cell self-renewal.</a> Stem Cells. 24 (6), 1505-1505 (2006).</li><li>Costoya, J. A., Hobbs, R. M., Barna, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Essential+role+of+Plzf+in+maintenance+of+spermatogonial+stem+cells.">Essential role of Plzf in maintenance of spermatogonial stem cells.</a> Nature Genetics. 36 (6), 653-653 (2004).</li><li>Morrow, C. M., Hostetler, C. E., Griswold, M. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+cold+storage+and+cryopreservation+of+immature+non-human+primate+testicular+tissue+on+spermatogonial+stem+cell+potential+in+xenografts.">Effect of cold storage and cryopreservation of immature non-human primate testicular tissue on spermatogonial stem cell potential in xenografts.</a> Human Reproduction. 22 (4), 1060-1060 (2007).</li><li>Rathi, R., Zeng, W., Megee, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maturation+of+testicular+tissue+from+infant+monkeys+after+xenografting+into+mice.">Maturation of testicular tissue from infant monkeys after xenografting into mice.</a> Endocrinology. 149 (10), 5288-5288 (2008).</li><li>Honaramooz, A., Li, M. W., Penedo, M. 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1. Germ celltransplantation Germ celltransplantation tillhandahåller den enda funktionell-analysen för otvetydig bekräftelse av närvaron av spermatogonial stamceller (SSC: er) i en cellpopulation. Endast SSC kan hem till och kolonisera SSC nisch vid basalmembranet och initiera donator-derived spermatogenesen. Könscell transplantation gjort det möjligt att studera och manipulera SSC på ett aldrig tidigare skådat sätt. Tekniken har använts för att framställa transgena djur genom gene...

<table border="1"><tbody><tr><td> Namnet på den reagenset </td><td> Bolaget </td><td> Katalognummer </td></tr><tr><td> Kollagenas (typ IV) </td><td> Sigma </td><td> C5138 </td></tr><tr><td> Trypsin-EDTA </td><td> Invitrogen </td><td> 25200-056 </td></tr><tr><td> DNasI </td><td> Sigma </td><td> DN25 </td></tr><tr><td> DMEM- </td><td> Invitrogen </td><td> 31053-028 </td></tr><tr><td> Trypanblått fläcka </td><td> Invitrogen </td><td> 15250-061 </td></tr><tr><td> Nylon mesh cellfilter </td><td> BD Biosciences </td><td> 352.340 (40 pm) 352.350 (70μm) </td></tr><tr><td> Busulfan </td><td> Sigma </td><td> B2635 </td></tr><tr><td> Thin-Wall glaskapillärer </td><td> World Precision Instrument </td><td> TW 100-3 </td></tr><tr><td> BD Intramedic plyethylene slang (PE100) </td><td> BD </td><td> CA-63018-725 </td></tr><tr><td> Ethicon 6-0 silkesutur </td><td> Ethicon </td><td> 706G </td></tr><tr><td> Sårklämmor </td><td> BD </td><td> 427.631 </td></tr><tr><td> Sigmacote </td><td> Sigma </td><td> SL2 </td></tr><tr><td> X-gal- </td><td> Sigma </td><td> B4252 </td></tr><tr><td> Kalium-Ferrocyanid </td><td> Sigma </td><td> P9387 </td></tr><tr><td> Kalium-Ferricyanid </td><td> Sigma </td><td> P3667 </td></tr><tr><td> magnesiumklorid </td><td> Sigma </td><td> 208.337 </td></tr><tr><td> natriumdeoxikolat </td><td> Sigma </td><td> D6750 </td></tr><tr><td> N, N-Dimetylformamid </td><td> Sigma </td><td> D4551 </td></tr><tr><td> Igepal CA-630 </td><td> Sigma </td><td> 18.896 </td></tr></tbody></table>

germ cell transplantation and testis tissue xenografting in mice

Könscell transplantation har utvecklats av Dr Ralph Brinster och kollegor vid University of Pennsylvania i 1994 1,2. Dessa banbrytande studier visade att mikroinjektion av könsceller från bördiga donatormöss i sädeskanalerna av alla infertila mottagande möss resulterar i givaren härlett spermatogenes och spermieproduktion av mottagaren djuret 2. Användningen av donatordjur bär den bakteriella β-galaktosidasgenen möjliggjorde identifiering av givare härrörande spermatogenes och överföring av givare haplotypen till avkomman med mottagande djur 1. Helt överraskande, efter transplantation in i lumen av sädeskanalernas tubuli, voro transplanterade könscellerna kunna röra sig från den luminala avdelningen till den källaren membranet där spermatogonier är belägna 3. Det är allmänt godtas att förordningen enbart SSC: er är i stånd att kolonisera den nisch-och återupprätta spermatogenesen hos de mottagande testikel. Därför könscelltransplantation ger ett funktionellt synsätt för att studera den nisch stamceller i testiklarna och för att karakterisera förmodade spermatogonial stamceller. Till datum, är grodden celltransplantation användas för att belysa grundläggande biologi stamcell, att producera transgena djur igenom genetisk manipulering av könsceller före transplantation 4,5, för att studera Sertolicellen-grodden cell-interaktion 6,7, Rymdbolaget homing och kolonisering 3, 8, samt SSC självförnyelse och differentiering 9,10. Könscell transplantation är också möjligt i stora arter 11. I dessa är de huvudsakliga användningsområdena är bevarandet av fertilitet, spridning av elit genetik i djurpopulationer, och generering av transgena djur som studiet av spermatogenes och SSC biologi med denna teknik är logistiskt svårare och dyrare än hos gnagare. Transplantation av könsceller från stora arter i sädeskanalerna av möss leder till colonization av donerade celler och spermatogonial expansion, men inte i sin fulla differentiering förmodligen på grund av inkompatibilitet mellan mottagaren somatiska cellen fack med könscellerna från fylogenetiskt avlägsna arter 12. Ett alternativt tillvägagångssätt är transplantation av könsceller från stora arter tillsammans med sin omgivning somatisk avdelning. Vi rapporterade först i 2002, att små fragment av testiklarna vävnad från omogna män transplanterade under dorsal huden immundefekta möss kan överleva och genomgå en fullständig utveckling med produktion av befruktning kompetent sperma 13. Sedan dess har testiklarna vävnad xenografting har visat sig vara framgångsrik i många arter och framträdde som ett värdefullt alternativ för att studera testiklar utveckling och spermatogenes av stora djur i möss 14.

Watch this Scientific Journal Video about Germ Cell Transplantation and Testis Tissue Xenografting in Mice at JoVE.com

Germ Cell Transplantation and Testis Tissue Xenografting in Mice

Protokoll för könsceller transplantation och testiklar xenografting vävnad beskrivs. Könsceller transplantation resultat i givar-derived spermatogenes i mottagarländerna testiklar och representerar en funktionell beredning analys för identifiering av spermatogonial stamceller (SSC). Testikel vävnaden xenografting återger testiklar utveckling och spermatogenes av olika givare arter i mottagande möss.

Könscell Transplantation och testiklar Tissue Xenografting hos möss

Germ cell transplantation was developed by Dr. Ralph Brinster and colleagues at the University of Pennsylvania in 19941,2. These ground-breaking studies ...

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