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

从作者实验室的工作已经支持农业部/ CSREES / NRICGP（2007-35203-18213）; NIH / NCRR（R01 RR17359-06），美国国立卫生研究院/ NIEHS的（1 R21的ES014856-01A2）和艾伯塔创新 - 健康解决方案。

A部分胚芽细胞移植在小鼠 1。受体小鼠的制备<ol><li>受助人应是免疫耐受（无论是捐助者或免疫缺陷的基因相匹配的）捐助睾丸细胞。 </li><li>收件人应当要么自然没有精子（ 如 W / 宽 V小鼠）或内源性的生殖细胞耗尽。可以通过辐射或化疗药物，如马利兰生殖细胞耗竭。在这个协议中，我们描述的准备与马利兰受体小鼠的方法。 </li><li>对待受助人的年龄...

<ol>
	<li>Brinster, R. L., Avarbock, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Germline+transmission+of+donor+haplotype+following+spermatogonial+transplantation.">Germline transmission of donor haplotype following spermatogonial transplantation.</a> Proceedings of the National Academy of Sciences of the United States of America. 91 (24), 11303-11303 (1994).</li><li>Brinster, R. L., Zimmermann, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spermatogenesis+following+male+germ-cell+transplantation.">Spermatogenesis following male germ-cell transplantation.</a> Proceedings of the National Academy of Sciences of the United States of America. 91 (24), 11298-11298 (1994).</li><li>Nagano, M., 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=Pattern+and+kinetics+of+mouse+donor+spermatogonial+stem+cell+colonization+in+recipient+testes.">Pattern and kinetics of mouse donor spermatogonial stem cell colonization in recipient testes.</a> Biology of Reproduction. 60 (6), 1429-1429 (1999).</li><li>Nagano, M., Brinster, C. J., Orwig, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+mice+produced+by+retroviral+transduction+of+male+germ-line+stem+cells.">Transgenic mice produced by retroviral transduction of male germ-line stem cells.</a> Proceedings of the National Academy of Sciences of the United States of America. 98 (23), 13090-13090 (2001).</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=Efficient+generation+of+transgenic+rats+through+the+male+germline+using+lentiviral+transduction+and+transplantation+of+spermatogonial+stem+cells.">Efficient generation of transgenic rats through the male germline using lentiviral transduction and transplantation of spermatogonial stem cells.</a> Journal of Andrology. 28 (2), 353-353 (2007).</li><li>Hess, R. A., Cooke, P. S., Hofmann, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanistic+insights+into+the+regulation+of+the+spermatogonial+stem+cell+niche.">Mechanistic insights into the regulation of the spermatogonial stem cell niche.</a> Cell Cycle. 5 (11), 1164-1164 (2006).</li><li>Oatley, M. J., Racicot, 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=Sertoli+cells+dictate+spermatogonial+stem+cell+niches+in+the+mouse+testis.">Sertoli cells dictate spermatogonial stem cell niches in the mouse testis.</a> Biology of Reproduction. 84 (4), 639-639 (2011).</li><li>Nagano, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Homing+efficiency+and+proliferation+kinetics+of+male+germ+line+stem+cells+following+transplantation+in+mice.">Homing efficiency and proliferation kinetics of male germ line stem cells following transplantation in mice.</a> Biology of Reproduction. 69 (2), 701-701 (2003).</li><li>Kubota, H., 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=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. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ETV5+is+required+for+continuous+spermatogenesis+in+adult+mice+and+may+mediate+blood+testes+barrier+function+and+testicular+immune+privilege.">ETV5 is required for continuous spermatogenesis in adult mice and may mediate blood testes barrier function and testicular immune privilege.</a> Annals of the New York Academy of Sciences. 1120, 144-144 (2007).</li><li>Zeng, W., Snedaker, A. K., 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=Preservation+and+transplantation+of+porcine+testis+tissue.">Preservation and transplantation of porcine testis tissue.</a> Reproduction, Fertility and Development. 21 (3), 489-489 (2009).</li><li>Jahnukainen, K., Ehmcke, J., Hergenrother, S. 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. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+maturation+of+primate+testis+by+xenografting+into+mice.">Accelerated maturation of primate testis by xenografting into mice.</a> Biology of Reproduction. 70 (5), 1500-1500 (2004).</li></ol>

1。生殖细胞移植生殖细胞移植提供了明确确认的精原干细胞存在于细胞的人口（SSCS）的唯一功能检测。只有共享服务中心可以回家和拓殖在基底膜SSC的利基，并发起捐助源性精子。生殖细胞移植使人们有可能以前所未有的方式SSC的研究和操纵。该技术已用于生产转基因动物通过SSCS 4的基因操纵澄清定植3,8南南合作的模式，效率和动力学研究信号通路调节SSC的?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>胶原酶（IV型） </td><td>西格玛</td><td> C5138 </td></tr><tr><td>胰蛋白酶EDTA </td><td> Invitrogen公司</td><td> 25200-056 </td></tr><tr><td> DNaseI </td><td>西格玛</td><td> DN25的</td></tr><tr><td> DMEM培养液</td><td> Invitrogen公司</td><td> 31053-028 </td></tr><tr><td>台盼蓝染色</td><td> Invitrogen公司</td><td> 15250-061 </td></tr><tr><td>尼龙网状细胞过滤</td><td> BD公司</td><td> 352340（40微米） 352350（为70μm） </td></tr><tr><td>马利兰</td><td>西格玛</td><td>具推拔孔</td></tr><tr><td>薄壁玻璃毛细管</td><td>世界精密仪器</td><td>台湾100-3 </td></tr><tr><td>的屋宇署intramedic plyethylene管材（PE100的） </td><td>屋宇署</td><td> CA-63018-725 </td></tr><tr><td>爱惜康6-0丝线缝合</td><td>爱惜康</td><td> 706G </td></tr><tr><td>伤口剪辑</td><td>屋宇署</td><td> 427631 </td></tr><tr><td> Sigmacote </td><td>西格玛</td><td> SL2的</td></tr><tr><td> X-gal的</td><td>西格玛</td><td> B4252 </td></tr><tr><td>亚铁氰化钾</td><td>西格玛</td><td> P9387 </td></tr><tr><td>铁氰化钾</td><td>西格玛</td><td> P3667 </td></tr><tr><td>氯化镁</td><td>西格玛</td><td> 208337 </td></tr><tr><td>去氧胆酸钠</td><td>西格玛</td><td> D6750 </td></tr><tr><td> ，N，N-二甲基甲酰胺</td><td>西格玛</td><td> D4551 </td></tr><tr><td> IGEPAL的CA-630 </td><td>西格玛</td><td> 18896 </td></tr></tbody></table>

germ cell transplantation and testis tissue xenografting in mice

生殖细胞移植是由拉尔夫Brinster博士和他的同事在美国宾夕法尼亚大学在1994年1,2。这些开创性的研究表明，捐助衍生的精子和精子生产由收件人动物2到不孕受体小鼠的结果曲细精管生精细胞注射从肥沃的供体小鼠。的使用捐赠受援国动物携带细菌β-半乳糖苷酶基因，鉴定捐助衍生的精子和捐赠单体的传输后代男性。令人惊讶的是，进入曲细精管的管腔移植后，移植的生殖细胞能够从管腔车厢移动其中精原细胞位于基底膜。它被普遍接受，唯一的SSC能够殖民的利基，并在收件人睾丸重新建立精子。因此，生殖细胞移植提供了一个功能的方法来研究在睾丸干细胞小生境的特点推测的精原干细胞。到今天为止，生殖细胞移植用于澄清基本的干细胞生物学，生殖细胞的遗传操作，通过生产转基因动物移植4,5之前，研究Sertoli细胞的生殖细胞相互作用6,7，SSC的归巢和定植3， 8，以及SSC的自我更新和分化9,10。 11大物种生殖细胞移植也是可行的。主要应用在这些保存生育能力，传播精英在动物种群遗传学，转基因动物精子和SSC生物学研究这一技术是后勤更加困难和昂贵的，比在啮齿类动物中的一代。从大型物种的生殖细胞移植到小鼠结果在colonizat曲细精管离子的供体细胞和精原干细胞扩张，但没有充分分化，大概是由于收件人的体细胞与生殖细胞系统发育遥远的物种12间不相容。另一种方法是从大型物种与周围的体车厢的生殖细胞的移植。我们首次报道于2002年，即从下背部皮肤移植免疫缺陷小鼠的不成熟男性睾丸组织的小碎片能够生存和接受的全面发展与生产的施肥主管精子13。此后睾丸组织异种移植已被证明是成功在许多物种中，并作为一种有价值的替代研究小鼠14大型动物的睾丸发育和精子出现。

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

生殖细胞移植和睾丸组织异种移植的协议描述。在捐助源性受援国睾丸精子的生殖细胞移植的结果和鉴定精原干细胞（SSCS）为代表的功能重建法。睾丸组织异种移植在受体小鼠睾丸发育和精子各捐助国物种再现。

生殖细胞的移植和睾丸组织在小鼠异种移植

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

Research

JoVE Journal

Biology

Isolation and Transplantation of Hematopoietic Stem Cells (HSCs)

Isolation and Transplantation of Hematopoietic Stem Cells HSCs

分离和造血干细胞移植（造血干细胞）

科研

生物学

Transplantation of Whole Kidney Marrow in Adult Zebrafish

在成年斑马鱼的全肾骨髓移植

Hematopoietic stem cells (HSC) are a rare population of pluripotent cells that maintain all the differentiated blood lineages throughout the life of an ...

Heterotopic Heart Transplantation in Mice

小鼠异位心脏移植

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

Small Bowel Transplantation In Mice

小鼠小肠移植

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive ...

Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin

肌动蛋白共沉淀测定;​​ F -肌动蛋白结合蛋白分析

The actin cytoskeleton within the cell is a network of actin filaments that allows the movement of cells and cellular processes, and that generates ...

Generation of Human CD40-activated B cells

人CD40激活的B细胞的生成

CD40-activated B cells (CD40-B cells) have been identified as an alternative source of immuno-stimulatory antigen-presenting cells (APC) for cancer ...

Tissue Targeted Embryonic Chimeras: Zebrafish Gastrula Cell Transplantation

组织有针对性的胚胎嵌合体：斑马鱼原肠胚细胞移植

Certain fundamental questions in the field of developmental biology can only be answered when cells are placed in novel environments or when small groups ...

Heterotopic and Orthotopic Tracheal Transplantation in Mice used as Models to Study the Development of  Obliterative Airway Disease

异位和原位气管移植小鼠模型研究闭塞性气道疾病的发展

Obliterative airway disease (OAD) is the major complication after lung transplantations that limits long term survival (1-7).
To study the ...

Planarian Immobilization, Partial Irradiation, and Tissue Transplantation

涡固定，局部照射，组织移植

The planarian, a freshwater flatworm, has proven to be a powerful system for dissecting metazoan regeneration and stem cell biology1,2. Planarian ...

Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations

在整个细胞群在单细胞水平上研究细胞周期蛋白B的蛋白水解

Equal distribution of chromosomes between the two daughter cells during cell division is a prerequisite for guaranteeing genetic stability 1. Inaccuracies ...