Name: parabiosis in mice: a detailed protocol
Uploaded: 2013-10-06T21:30:00
Description: Watch this Scientific Journal Video about Parabiosis in Mice: A Detailed Protocol at JoVE.com

作者要感谢阿德里恩莫斯利和Libuse耶扎贝克（斯坦福大学），用于与手术技术援助。

所有的动物研究，根据加州大学洛杉矶分校的动物护理和使用委员会实验动物的护理和使用的指导方针和卫生指南的国家机构进行。下面描述的步骤的持续时间是从开始约45-60分钟来结束。 1。手术野的制备<ol><li>在一个干净的动物手术室执行程序。 </li><li>设备：异氟醚蒸发器，盖默牛逼泵用加热垫。 </li><li>无菌工具：两弯钳，细剪刀，持针器。 </li><li>无菌手套必须在整个...

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	<li>Bert, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exp&#233;riences+et+consid&#233;rations+sur+la+greffe+animale.">Exp&#233;riences et consid&#233;rations sur la greffe animale.</a> Journal de l'Anatomie et de la Physiologie. 1, 69-87 .</li><li>Bunster, E., Meyer, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+methods+of+parabiosis.">Improved methods of parabiosis.</a> Anat. Rec. 57, 339-380 (1933).</li><li>Waskow, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+parabiotic+mice+for+the+study+of+DC+and+DC+precursor+circulation.">Generation of parabiotic mice for the study of DC and DC precursor circulation.</a> Methods Mol. Biol. 595, 413-428 (2010).</li><li>Finerty, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parabiosis+in+physiological+studies.">Parabiosis in physiological studies.</a> Physiol. Rev. 32, 277-302 (1952).</li><li>Aicher, A., Heeschen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonbone+marrow-derived+endothelial+progenitor+cells:+what+is+their+exact+location.">Nonbone marrow-derived endothelial progenitor cells: what is their exact location.</a> Circ. Res. 101, e102 (2007).</li><li>Abe, S., Boyer, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cells+derived+from+the+circulation+contribute+to+the+repair+of+lung+injury.">Cells derived from the circulation contribute to the repair of lung injury.</a> Am. J. Respir. Crit. Care Med. 170, 1158-1163 (2004).</li><li>Donskoy, E., Goldschneider, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thymocytopoiesis+is+maintained+by+blood-borne+precursors+throughout+postnatal+life.+A+study+in+parabiotic+mice.">Thymocytopoiesis is maintained by blood-borne precursors throughout postnatal life. A study in parabiotic mice.</a> J. Immunol. 148, 1604-1612 (1992).</li><li>Powell, A. E., Anderson, E. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fusion+between+Intestinal+epithelial+cells+and+macrophages+in+a+cancer+context+results+in+nuclear+reprogramming.">Fusion between Intestinal epithelial cells and macrophages in a cancer context results in nuclear reprogramming.</a> Cancer Res. 71, 1497-1505 (2011).</li><li>Duyverman, A. M., Kohno, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+transient+parabiosis+skin+transplantation+model+in+mice.">A transient parabiosis skin transplantation model in mice.</a> Nat. Protoc. 7, 763-770 (2012).</li><li>Ajami, B., Bennett, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+self-renewal+can+sustain+CNS+microglia+maintenance+and+function+throughout+adult+life.">Local self-renewal can sustain CNS microglia maintenance and function throughout adult life.</a> Nat. Neurosci. 10, 1538-1543 (2007).</li><li>Weissman, I. L., Jerabek, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tolerance+and+the+H-Y+antigen:+Requirement+for+male+T+cells,+but+not+B+cells,+to+induce+tolerance+in+neonatal+female+mice.">Tolerance and the H-Y antigen: Requirement for male T cells, but not B cells, to induce tolerance in neonatal female mice.</a> Transplantation. 37, 3-6 (1984).</li></ol>

这里讨论的联体的方法呈现在低死亡率最小的技术难点和结果。膝关节和肘关节的附件是一个显著改善本斯特和迈耶技术。但是，该过程仍侵入从而维持无菌条件下整个手术是势在必行。以进一步防止外科手术部位的感染，这是很重要的parabiosed动物接受抗生素的组合，进行定期监测。为确保坚定的支持，防止疼痛，连接肘部和膝盖缝合应围绕关节，而不是通过组织。 作为替?...

联体，手术加盟的两种生物，于1864年首次描述了保罗·伯特，以此来建立一个模型来研究共享循环系统和由两只老鼠1的皮肤和肌肉壁的加盟。联体促进微血管的形成在3发炎的部位，并有多个应用程序在生理的研究，如脑下垂体和性腺与肾脏高血压4的作用之间的荷尔蒙沟通。它得到了进一步的来探讨祖细胞的招募和整合新生血管5，造血干细胞6的迁移和淋巴细胞贩运7，以及在肿瘤转移8,9循环炎性细胞或干细胞的作用和动力学，和神经退行性疾病10。 联体谎言中的一个显著优势在该合作的动物有着共同的循环抗原，使细胞迁移和血管新生，而不会触发免疫反应。重要的是，威斯曼等人已经显示雄性和雌性小鼠之间的联体不会导致形成抗HY的抗体11。 在由Paul伯特描述的原始协议的2只动物经皮肤和肌肉壁1的连接接合在一起。此方法然而，引起显著菌株对动物和由于伤口的感染导致高死亡率。自那时以来，联体技术已经修订了几组最最主要的是在1933年2提出的本斯特和迈耶的协议。他们的方法包括加入肩胛骨关节，体腔和皮肤，允许更好的支持和痛苦少的动物。与此同时，新的方法导致了最小的术后护理和显着地降低死亡率。本文所描述的协议是本斯特和迈耶技术，是微创的，并允许更牢固接合的变形。即，将小鼠通过肘和膝盖关节以及皮肤相连。皮肤的这种连接可以防止分机，因此导致更少的痛苦和并发症。在这里，我们描述了野生型（WT）成年小鼠的组成型表达GFP的小鼠的加盟。我们发现，手术后两周我们可以达到血液嵌合体的50％证明了这种外科手术的功效，以创建一个共享的循环系统。

<table border="1">
	<tbody>
		<tr>
			<td>Name of equipment</td>
			<td>Company</td>
			<td>Catalog number</td>
		</tr>
		<tr>
			<td>Isoflurane-Phoenix</td>
			<td>Clipper</td>
			<td>NDC 57319-559-06</td>
		</tr>
		<tr>
			<td>Posi-Seal Induction Chamber</td>
			<td>Molecular Imaging Products</td>
			<td>AS-01-0530-SM</td>
		</tr>
		<tr>
			<td>Portable Anesthesia System</td>
			<td>Molecular Imaging Products</td>
			<td>AS-01-0007</td>
		</tr>
		<tr>
			<td>Gaymer T Pump</td>
			<td>Gaymar Industries, Inc</td>
			<td>TP650</td>
		</tr>
		<tr>
			<td>Warming Blanket (Heating pad)</td>
			<td>Kent Scientific Corp</td>
			<td>TP-22G</td>
		</tr>
		<tr>
			<td>Curved forceps</td>
			<td>Roboz</td>
			<td>RS-5101</td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Fine Science Tools (FST)</td>
			<td>FST 14063-09</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>FST</td>
			<td>FST 12501-13</td>
		</tr>
		<tr>
			<td>Electrical shaver</td>
			<td>Oster</td>
			<td>Golden A5</td>
		</tr>
	</tbody>
</table>

parabiosis in mice: a detailed protocol

联体是两种生物体，允许共享的血液循环的外科结合。将两个动物的皮肤促进形成微脉管系统中的炎症部位。连体合作伙伴分享他们的循环抗原，因此是免费的不良免疫反应。首先由保罗·伯特在1864年1所述，联体手术是通过提炼和本斯特迈耶于1933年，以改善动物生存2。在当前的协议中，2只小鼠手术加入以下的本斯特和迈耶技术的一种改进。动物通过肘部和膝关节后面附着在皮肤允许，防止紧张的缝合皮肤坚定支持的连接。在此，我们详细描述了连体加入一个无处不在的绿色荧光蛋白表达小鼠与野生型（WT）小鼠。手术后两周，对被分离和GFP阳性细胞可以通过流式细胞术分析来检测血液中的circulat离子的WT鼠标。血液嵌合允许一个检查从在另一种动物的循环细胞的贡献。

的两个生物体联体的预期结果是每只动物的循环系统的一个共同的血液循环（ 图2）的相等的贡献。人们可以很容易验证parabiosed WT和GFP阳性小鼠经流式细胞仪分析血液的成功平衡。这里，从两个parabionts的尾部，得到在2周时，手术后静脉采血并进行分级分离为外周血细胞（排除红细胞）。分级分离的造血来源的细胞级分，随后用流式细胞仪对GFP阳性和WT细胞的存在进行分析。与以前的?...

Watch this Scientific Journal Video about Parabiosis in Mice: A Detailed Protocol at JoVE.com

Parabiosis in Mice: A Detailed Protocol

联体小鼠：一个详细的协议

联体接合的两个生物体导致的共享循环系统的发展。在这个协议中，我们描述了手术步骤，以形成一个野生型小鼠和组成型表达GFP的小鼠之间的联体连接。

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of ...

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