Name: formation of human prostate epithelium using tissue recombination of rodent urogenital sinus mesenchyme and human stem cells
Uploaded: 2013-06-22T14:00:00
Description: Watch this Scientific Journal Video about Formation of Human Prostate Epithelium Using Tissue Recombination of Rodent Urogenital Sinus Mesenchyme and Human Stem Cells at JoVE.com

我们希望大学由Arieh已Shalhav博士率领芝加哥科泌尿外科，泌尿研究嘉莉博士RINKER谢弗主任的支持表示感谢。我们还想大学芝加哥综合癌症中心（UCCCC）LE BEAU米歇尔博士领导的支持表示感谢。我们还与，感谢专家技术援助的人体组织资源中心的核心设施马克林根博士的带领下，和莱斯利·马丁和玛丽·乔·菲克特援助。我们也感谢李特丽免疫组化核心设备运行。这项工作是由芝加哥外科部泌尿外科，该​​科大学，美国癌症协会的研究资助机构（ACS-IRG IRG-58-004）;癌症中心的支持格兰特（P30 CA14599）;布林森基金会;阿尔文鲍姆家庭基金;大学芝加哥癌症研究基金会的董事会; S.克雷格尔由霍华德休斯医学研究所的支持：到毕业的医学研究员髋部（56006772）和癌症生物学训练津贴（T32-CA09594）。最后，我们要感谢罗伯特·克拉克，和弥敦道Stadick博士VenkateshKrishnan，都为他们的批判性评价的稿件。

这项研究进行了严格按照美国国立卫生研究院的实验室动物护理和使用指南的建议。该协议被批准大学芝加哥机构动物护理和使用委员会（IACUC，协议号码72066和72231）。所有的手术麻醉下进行，所有的努力都是为了最大限度地减少痛苦。收购WiCell（麦迪逊，威斯康星州）和培养人类胚胎干细胞系WA01（H1; NIH注册＃0043）使用独立的馈线协议使用mTeSR1媒体（干细胞技术;温哥华，不列颠哥伦比亚省）。...

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组织重组使用UGSM是一个非常有用的技术来探讨前列腺，导致前列腺癌症的发生的分子事件的发展。已被用于众多应用在前列腺研究中的感应电势UGSM，其中包括提高肿瘤的前列腺细胞系和肿瘤研究上皮间质的相互作用，并形成跨物种前列腺重组体。7,17-20适当的准备UGSM然而，实验成功，将导致污染啮齿动物上皮腺腔形成，可以混淆结果是至关重要的。因此，分离的UGSM从UGSE中（步骤2.4）是?...

为更好的人体模型系统对前列腺癌有一个巨大的需要。尤其是，相关的人体模型系统的正常，非恶性前列腺组织可以操纵基因直接辨别特定基因的作用在前列腺癌的启动将是令人难以置信的信息。基因组时代的来临已经确定了许多基因可能在癌症的形成有一定的作用。然而，缺乏人体实验模型系统严重损害我们的能力，功能测试和表征候选前列腺癌易感基因。一个理想的模型系统，将有利于迅速和更迅速的癌症易感基因组合与适当的转基因啮齿动物模型系统的功能分析。此外，这种人体模型系统将使前列腺癌变的信号机制向发现和验证新疗法的分子特征预防前列腺癌的形成。 人胚胎干细胞（人胚胎干细胞）是能够形成作为异种移植的人类前列腺组织。泰勒等人在2006年报道，人类胚胎干细胞可以诱导在体内与啮齿动物的泌尿生殖窦间充质（UGSM）在8-12周的时间段之内重新组合以形成前列腺上皮细胞。1这些研究是基于以前的工作库尼亚实验室表明，啮齿类动物胚胎UGSM可以促进前列腺分化的干细胞和胚胎在体内的上皮细胞群。2,3前列腺癌的发展从一个胚胎原基称为泌尿生殖窦（UGS），胚胎17天前（E17鼠标一天E18在大鼠）UGS可以删除，并在物理上分为上皮（UGSE）和间充质（UGSM）4本组织重组方法已明显使我们进一步了解前列腺的发育和癌变，特别是光年生长因子和激素信号转导途径和前列腺基质和上皮细胞的分子之间的关系。5-8此方法涉及体外的UGSM组合与干细胞或上皮细胞中的相同或不同的种属，这些细胞/组织的重组体注入和生长经过一段时间的在体内的生长和小鼠异种移植物于主机。4,9，所述植入物包含嵌入在基质组织的前列腺上皮腺状结构。进一步的染色，也可进行，以确定这样的结构是否是真正的前列腺和人类起源。10,11

<table border="1">
 <tbody>
 <tr>
 <td>Name</td>
 <td>Company</td>
 <td>Catalogue Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Hank's Balanced Salt Solution (HBSS)</td>
 <td>GIBCO</td>
 <td>14170</td>
 <td></td>
 </tr>
 <tr>
 <td>DMEM/F12</td>
 <td>GIBCO</td>
 <td>11330</td>
 <td></td>
 </tr>
 <tr>
 <td>R1881</td>
 <td>Sigma</td>
 <td>965-93-5</td>
 <td>Mix to 1 ug/ml in Ethanol (1,000x stock)</td>
 </tr>
 <tr>
 <td>NEAA</td>
 <td>GIBCO</td>
 <td>11140</td>
 <td></td>
 </tr>
 <tr>
 <td>Pen-Strep Solution</td>
 <td>GIBCO</td>
 <td>15070</td>
 <td>100x stock</td>
 </tr>
 <tr>
 <td>Matrigel</td>
 <td>BD Biosciences</td>
 <td>354230</td>
 <td></td>
 </tr>
 <tr>
 <td>KETASET (ketamine hydrochloride) </td>
 <td>Fort Dodge Animal Health</td>
 <td>NDC 0856-2013-01</td>
 <td>100 mg/ml; dilute 1:10 in sterile saline </td>
 </tr>
 <tr>
 <td>AnaSed (xylazine) </td>
 <td>VET-A-MIX, Inc.</td>
 <td>NADA 139-236</td>
 <td>20 mg/ml; dilute 1:10 in sterile saline</td>
 </tr>
 <tr>
 <td>Trypsin</td>
 <td>BD Biosciences</td>
 <td>215240</td>
 <td></td>
 </tr>
 <tr>
 <td>Collagenase </td>
 <td>Sigma</td>
 <td>C2014</td>
 <td></td>
 </tr>
 <tr>
 <td>Ketoprofen </td>
 <td>Fort Dodge</td>
 <td>NDC 0856-4396-01</td>
 <td>100 mg/ml; dilute 1:1,000 in sterile saline</td>
 </tr>
 <tr>
 <td>Altalube eye ointment </td>
 <td>Altaire Pharmaceuticals, Inc.</td>
 <td>NLC 56641-19850</td>
 <td></td>
 </tr>
 <tr>
 <td>Leica MZ16 F Stereomicroscope</td>
 <td>Leica</td>
 <td></td>
 <td>Any good dissecting scope can be used.</td>
 </tr>
 <tr>
 <td>Vannas spring scissors</td>
 <td>Fine Science Tools</td>
 <td>15001-08</td>
 <td></td>
 </tr>
 <tr>
 <td>Syringe</td>
 <td>Hamilton</td>
 <td>84855</td>
 <td></td>
 </tr>
 <tr>
 <td>Hamilton Needle, Small RN, 28 gauge, 0.5inches, Point Style #3 (Blunt)</td>
 <td>Hamilton</td>
 <td>7803-02 </td>
 <td>Custom Needle </td>
 </tr>
 <tr>
 <td>Ethanol Prep Pads</td>
 <td>Fisher Scientific</td>
 <td>06-669-62 </td>
 <td></td>
 </tr>
 <tr>
 <td>Sterile Gauze Pads</td>
 <td>Fisher Scientific</td>
 <td>22-415-469 </td>
 <td></td>
 </tr>
 <tr>
 <td>Ethicon Vicryl Suture (4-0 FS-2)</td>
 <td>MedVet International</td>
 <td>J392H </td>
 <td>Needle-in, dissolvable suture </td>
 </tr>
 <tr>
 <td>Autoclip 9 mm Wound Clips</td>
 <td>Becton Dickenson</td>
 <td>427631</td>
 <td></td>
 </tr>
 <tr>
 <td>PVP Iodine Prep Pads</td>
 <td>Fisher Scientific</td>
 <td>06-669-98 </td>
 <td></td>
 </tr>
 <tr>
 <td>Dissector scissor with blunt end</td>
 <td>Fine Science Tools</td>
 <td>14072-10</td>
 <td></td>
 </tr>
 <tr>
 <td>Dumont fine tip forceps</td>
 <td>Fine Science Tools</td>
 <td>11252-50</td>
 <td></td>
 </tr>
 <tr>
 <td>Needle holder with Scissor</td>
 <td>Fine Science Tools</td>
 <td>12002-14</td>
 <td></td>
 </tr>
 </tbody>
</table>

formation of human prostate epithelium using tissue recombination of rodent urogenital sinus mesenchyme and human stem cells

人源性和激素天真的模型系统，这限制了我们的能力，了解相关前列腺疾病引发的遗传和分子事件的可用性，在前列腺癌的研究进展受到严重限制。对发展更好的模型系统研究人类前列腺癌，我们和其他人利用胚胎鼠的前列腺基质亲前列腺独特的感应电势，被称为泌尿生殖窦间充质（UGSM）。当重组的目标，如胚胎干细胞的多能性的细胞群，UGSM睾酮依赖性诱导正常人类前列腺上皮细胞的形成。这种人体模型系统可用于调查和实验测试候选前列腺癌易感基因的能力比较典型的啮齿动物转基因研究加快。由于人类胚胎干细胞（胚胎）可以在全光照培养转基因克诱导基因表达或siRNA击倒向量组织重组之前，这样的模式有利于测试基因功能的后果，或基因组合，这被认为是促进或防止癌变的。 然而，分离纯人群的UGSM细胞，该技术的挑战和学习往往需要与以前的专业知识的人亲自教导。此外，肾包膜下的细胞混合物接种的免疫功能低下的主机可以是技术上的挑战。这里，我们勾勒，说明适当的隔离的UGSM从啮齿动物胚胎组织混合物形成人类前列腺上皮和肾囊的着床。这样的方法，在目前的阶段，需要在体内异种移植的胚胎干细胞，未来的应用程序可能会包括在体外腺形成或使用诱导式多能性干细胞群（iPS细胞）。

泰勒等人的精彩的 ​​报告基础上，我们的实验室已研制出一种嫁接协议使用常用的H1（NIH-WA01指定，男性基因）人类胚胎干细胞系。1这行已经严格测试，质量控制和核型正常的13，当适当的培养，胚胎干细胞可以保持，扩大，并使用无饲养层培养方法（馈线系统通过干细胞技术市售温哥华BC），冷冻保存在一个未分化和多能状态。14我们协议采用结合与单细胞悬液E...

Watch this Scientific Journal Video about Formation of Human Prostate Epithelium Using Tissue Recombination of Rodent Urogenital Sinus Mesenchyme and Human Stem Cells at JoVE.com

Formation of Human Prostate Epithelium Using Tissue Recombination of Rodent Urogenital Sinus Mesenchyme and Human Stem Cells

为了解开最早的分子机制启动前列腺癌，新颖创新的人体模型系统和方法是迫切需要的。诱导多能干细胞群形成的人类前列腺上皮细胞的潜力预前列腺泌尿生殖窦间充质（UGSM）是一个强大的实验工具，在前列腺研究。

人类前列腺上皮细胞形成的使用鼠类泌尿生殖窦间质细胞和人类干细胞的组织重组

Progress in prostate cancer research is severely  limited by the availability of human-derived and hormone-na&iuml;ve model systems,  which limit our ...

Research

JoVE Journal

Medicine

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

在干细胞移植在体外模拟缺血/再灌注模型

Stem cell transplantation protocols are finding their way into clinical practice1,2,3. Getting better results, making the protocols more robust, and ...

科研

医学

Subretinal Injection of Gene Therapy Vectors and Stem Cells in the Perinatal Mouse Eye

基因治疗的载体和干细胞视网膜下注射到老鼠的眼睛

The loss of sight affects approximately 3.4 million people in the United States and is expected to increase in the upcoming years.1 Recently, gene therapy ...

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

人牙髓干细胞的分离，鉴定和比较性恒牙使用两种不同的方法，得出

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

人类前列腺癌转移的原位小鼠模型

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Isolation of Cancer Stem Cells From Human Prostate Cancer Samples

癌症干细胞从人类前列腺癌样本隔离

The cancer stem cell (CSC) model has been considerably revisited over the last two decades. During this time CSCs have been identified and directly ...

Enrichment for Chemoresistant Ovarian Cancer Stem Cells from Human Cell Lines

丰富的人力细胞系化疗耐药性卵巢癌干细胞

Cancer stem cells (CSCs) are defined as a subset of slow cycling and undifferentiated cells that divide asymmetrically to generate highly proliferative, ...

Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays

评价的人卵巢癌细胞干细胞特性使用多和单细胞为基础的化验球

Years of research indicates that ovarian cancers harbor a heterogeneous mixture of cells including a subpopulation of so-called &#8220;cancer stem ...

Laser-capture Microdissection of Human Prostatic Epithelium for RNA Analysis

人类前列腺上皮的激光捕获显微切割的RNA分析

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types.  Healthy prostate is comprised of ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

人前列腺癌的临床前原位小鼠模型

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Generation of a Humanized Mouse Liver Using Human Hepatic Stem Cells

利用人体肝干细胞人源化小鼠肝脏生成

A novel animal model involving chimeric mice with humanized livers established via human hepatocyte transplantation has been developed. These mice, in ...