Name: orthotopic implantation and peripheral immune cell monitoring in the ii-45 syngeneic rat mesothelioma model
Uploaded: 2015-10-02T14:00:00
Description: Watch this Scientific Journal Video about Orthotopic Implantation and Peripheral Immune Cell Monitoring in the II-45 Syngeneic Rat Mesothelioma Model at JoVE.com

The syngeneic rat mesothelioma II-45 cell line was a kind gift from A/Prof. Emanuela Felley-Bosco, Zurich University.

所有涉及动物的程序进行，根据实践中的动物的护理和使用用于科学目的的澳大利亚准则的建议。该协议为这项研究是经皇家北岸医院的动物护理和伦理委员会。女菲舍尔344大鼠（F344，150-200克）保持在卡恩斯设施，Kolling研究所标准条件（12小时亮/暗周期和自由获得食物和水）下。 注意：一个流程图，用于所有实验程序是在图1。 ...

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本文详述了胸膜间皮瘤大鼠的同源原位模型以及用于通过纵向血液取样监测疾病进展的简单方法的产生的方法。 在II-45模型通过将菲舍尔344大鼠石棉纤维13开发的。虽然这种曝光表示主机石棉免疫系统的相互作用的真实动态间皮瘤发病机制，它具有长的滞后时间（以年来产生），并可能是危险的，研究人员由于可能暴露于石棉纤维。 Ⅱ-45细胞的原位移植进入胸膜动物?...

恶性间皮瘤（MM）是一种侵袭性的恶性肿瘤其来自转化的细胞中的膜（间皮），该系肺和腹腔，心脏和内部繁殖器官和是肺腔或胸膜1,2的最常见的原发性肿瘤。暴露于石棉纤维占80％，所有的MM，虽然对石棉的使用禁令推出几十年前，在大多数西方国家，其社会各界的广泛使用已经留下了一个致命的遗产。据世界卫生组织估计，107,000名员工每年死于与石棉有关的疾病，与死亡率持续增加。一种新的非职业入射波也正在和有何时，在什么水平，这将达到峰值3知之甚少。 大多数人的MM被确诊晚了，当全身化疗代表的唯一可行的选择4之一。最effecti已经化疗（顺铂5培美曲塞在一起）当前的“护理标准”被认定在10年前。但是失败这种治疗是不可避免的，也没有成熟的第二行选项，使患者的一个严峻的预后只有12 个月2，中位生存期。因此，目前迫切未满足的需要更有效的治疗方法。尽管一批临床试验中的新疗法的检查没有导致的变化在实际中。这是部分由于低5％转移的临床前的结果，一般在异种移植小鼠模型中进行，给临床设定6-8。这类模型不忠实概括在非生理位置中发生的肿瘤微环境的复杂的方面，经常在没有免疫系统功能9。 同源原位模型创建比C一显著更现实的肿瘤环境ommonly使用皮下异种移植模型作为出现在正确的生理部位有一个完整的免疫系统10,11的肿瘤。的大鼠的尺寸较大增强其作为一种啮齿动物疾病模型的使用，尤其是在药物研究，其中串行抽血需要来评估治疗反应和毒性12。此外，在模型中，监测疾病进展是困难的，因为在肿瘤（如在胸膜腔）的位置，用在血液循环中发现的因素是非常有吸引力的，监测疾病进展的能力。使用免疫能干大鼠胸膜间​​皮瘤的同基因原位模型的生成进行说明。此外，一种简单的和相对非侵入性的方法，通过测量循环免疫细胞监测胸膜疾病进展还描述。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>EDTA Collection tube (0.5ml)</td>
			<td>Greiner Bio One GmbH</td>
			<td>450480</td>
			<td></td>
		</tr>
		<tr>
			<td>Rat T/B/NK Cell cocktail</td>
			<td>BD Pharmingen</td>
			<td>558509</td>
			<td>anti-Rat CD3&#160; APC (IgM clone 1F4), anti-Rat CD45RA Fitc (IgG1 clone OX-33), anti-Rat CD161a PE (IgG1 Clone 10/78)</td>
		</tr>
		<tr>
			<td>anti-RAT CD8a PE&#160;</td>
			<td>Biolegend</td>
			<td>200608</td>
			<td>(IgG1vClone G28)</td>
		</tr>
		<tr>
			<td>anti-Rat CD4 FITC (Domain 1)&#160;</td>
			<td>Biolegend</td>
			<td>203406</td>
			<td>(IgG1 Clone OX-38)</td>
		</tr>
		<tr>
			<td>anti-Rat CD45 PE/Cy7&#160;</td>
			<td>Biolegend</td>
			<td>202214</td>
			<td>(IgG1 Clone OX-1)</td>
		</tr>
		<tr>
			<td>TruCount&#8482; Tubes</td>
			<td>Becton Dickinson</td>
			<td>340334</td>
			<td>Box of 50 absolute counting tubes</td>
		</tr>
		<tr>
			<td>RPMI 1640 media</td>
			<td>Life Technologies</td>
			<td>11875-119</td>
			<td></td>
		</tr>
		<tr>
			<td>foetal bovine serum (FBS)</td>
			<td>Scientifix</td>
			<td>FBS500-S (lot# 010101-1)</td>
			<td></td>
		</tr>
		<tr>
			<td>trypsin-EDTA</td>
			<td>Life Technologies</td>
			<td>15400-054</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS tablets</td>
			<td>Medicago AB</td>
			<td>09-9400-100</td>
			<td></td>
		</tr>
		<tr>
			<td>23Gx1&#188; Needle</td>
			<td>Becton Dickinson</td>
			<td>302008</td>
			<td></td>
		</tr>
		<tr>
			<td>1mL Syringe</td>
			<td>Becton Dickinson</td>
			<td>302 100</td>
			<td></td>
		</tr>
		<tr>
			<td>Fischer 344 Rat</td>
			<td>Animal Resources Centre, Perth Australia</td>
			<td>F344</td>
			<td></td>
		</tr>
		<tr>
			<td>I.S.O (Isoflurane USP)</td>
			<td>Veterinary Companys Australia (VCA)&#160;</td>
			<td>B7058</td>
			<td></td>
		</tr>
		<tr>
			<td>II-45 Rat Mesothelioma line</td>
			<td>Zurich University</td>
			<td></td>
			<td>Note: The cell line was given as a gift and is not commercially available at the ATCC</td>
		</tr>
		<tr>
			<td>FACSCalibur&#8482; 4 colour</td>
			<td>Becton Dickinson</td>
			<td>342975</td>
			<td></td>
		</tr>
		<tr>
			<td>TRIS-HCL</td>
			<td>SIGMA</td>
			<td>T3253</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium Chloride</td>
			<td>SIGMA</td>
			<td>9718</td>
			<td></td>
		</tr>
		<tr>
			<td>Anaesthetic Machine (The stinger)</td>
			<td>Advanced Anaesthesia specialists</td>
			<td>#00449</td>
			<td></td>
		</tr>
	</tbody>
</table>

orthotopic implantation and peripheral immune cell monitoring in the ii-45 syngeneic rat mesothelioma model

在免疫为基础的治疗的癌症如疫苗和免疫检查点抑制剂，和在治疗响应肿瘤微环境中的作用的进一步了解感兴趣的巨大热潮，共同指向需要免疫能干正位模型的临床前试验这些新的治疗方法。本文演示了如何建立胸膜恶性间皮瘤原位免疫能力的动物模型。在原位模型监测疾病进展是由肿瘤的内部位置混淆。纵向监测疾病的进展及其对循环免疫细胞在这方面和癌症等大鼠模型的效果，单管流式细胞术检测只需要25全血中描述微升。这提供了七个免疫指标准确定量：总淋巴细胞，单核细胞和嗜中性粒细胞，以及T细胞亚群CD4和CD8，B细胞和自然杀伤细胞。不同的潜艇这些参数的ETS在不同的情况和模型有用，与中性粒细胞淋巴细胞比率具有用于监测疾病进程中的间皮瘤模型最大的效用。分析循环使用这种单管方法还可以协助监测响应免疫为基础的治疗和了解，导致治疗成功或失败的根本机制免疫细胞的水平。

在本文用于使用II-45细胞胸膜间皮瘤的原位模型​​的生成的方法，导致动物屈服于间皮瘤可再现的和快速的时间内，没有大鼠垂死由于植入方法。细胞植入的数量的滴定中确定1×10 3个细胞是需要的充分渗透模型（100％的植入）的最小数目。的不同数目的细胞植入到大鼠的改变疾病的时间过程中没有出现影响疾病的严重程度。动物接受1×10 3，1×10 4和5×10 5个细胞40天，30和...

Watch this Scientific Journal Video about Orthotopic Implantation and Peripheral Immune Cell Monitoring in the II-45 Syngeneic Rat Mesothelioma Model at JoVE.com

Orthotopic Implantation and Peripheral Immune Cell Monitoring in the II-45 Syngeneic Rat Mesothelioma Model

胸膜恶性间皮瘤由II-45间皮瘤细胞进入胸膜免疫活性大鼠腔植入的原位大鼠模型的产生提出。甲流式细胞法分析从25微升血样在这些动物7免疫细胞亚群也被描述。

在II-45同基因大鼠模型间皮瘤原位移植和外周免疫细胞监视

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用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...