Normal and malignant muscle cell transplantation into immune compromised adult zebrafish.
Normal and malignant muscle cell transplantation into immune compromised adult zebrafish.
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DOI：
10.3791/52597
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发表时间：
2014-12-26
期刊：
影响因子：
--
通讯作者：
Langenau DM
中科院分区：
文献类型：
--
作者：
Tenente IM;Tang Q;Moore JC;Langenau DM
斑马鱼已经成为评估发育、再生和癌症的有力工具。最近，同种异体移植细胞移植方案已经开发，允许正常和恶性细胞植入到辐射，同源，免疫受损的成年斑马鱼。这些模型与优化的细胞移植方案相结合，可以快速评估干细胞功能，损伤后再生和癌症。在这里，我们提出了一种方法，用于斑马鱼成年骨骼肌和胚胎横纹肌肉瘤（ERMS），一种儿科肉瘤，与胚胎肌肉共享功能，到免疫受损的成年rag 2 E450 fs纯合突变斑马鱼的细胞移植。重要的是，这些动物缺乏T细胞，并且B细胞功能降低，有利于来自无关供体动物的广泛组织的移植。我们优化的方案表明，荧光标记的肌肉细胞制剂从α-肌动蛋白RFP转基因斑马鱼植入到背部肌肉组织的rag 2纯合子突变鱼强劲。我们还证明了荧光转基因ERMS的植入，其中荧光仅限于基于分化状态的细胞。具体地，在AB-菌株myf 5-GFP; mylpfa-mCherry双转基因动物和注射到成年免疫受损鱼的腹膜中的肿瘤中产生ERMS。这些协议的效用扩展到广泛的正常和恶性供体细胞，可以植入背部肌肉组织或腹膜的成年斑马鱼的植入。
Zebrafish have become a powerful tool for assessing development, regeneration, and cancer. More recently, allograft cell transplantation protocols have been developed that permit engraftment of normal and malignant cells into irradiated, syngeneic, and immune compromised adult zebrafish. These models when coupled with optimized cell transplantation protocols allow for the rapid assessment of stem cell function, regeneration following injury, and cancer. Here, we present a method for cell transplantation of zebrafish adult skeletal muscle and embryonal rhabdomyosarcoma (ERMS), a pediatric sarcoma that shares features with embryonic muscle, into immune compromised adult rag2E450fs homozygous mutant zebrafish. Importantly, these animals lack T cells and have reduced B cell function, facilitating engraftment of a wide range of tissues from unrelated donor animals. Our optimized protocols show that fluorescently labeled muscle cell preparations from α-actin-RFP transgenic zebrafish engraft robustly when implanted into the dorsal musculature of rag2 homozygous mutant fish. We also demonstrate engraftment of fluorescent-transgenic ERMS where fluorescence is confined to cells based on differentiation status. Specifically, ERMS were created in AB-strain myf5-GFP; mylpfa-mCherry double transgenic animals and tumors injected into the peritoneum of adult immune compromised fish. The utility of these protocols extends to engraftment of a wide range of normal and malignant donor cells that can be implanted into dorsal musculature or peritoneum of adult zebrafish.
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影响因子：
50.3
作者：
Blackburn JS;Liu S;Wilder JL;Dobrinski KP;Lobbardi R;Moore FE;Martinez SA;Chen EY;Lee C;Langenau DM
通讯作者：
Langenau DM
DOI：
10.3791/50846
发表时间：
2014-04-08
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Motohashi N;Asakura Y;Asakura A
通讯作者：
Asakura A
影响因子：
50.3
作者：
Ignatius, Myron S.;Chen, Eleanor;Langenau, David M.
通讯作者：
Langenau, David M.
影响因子：
2.7
作者：
Higashijima, S;Okamoto, H;Eguchi, G
通讯作者：
Eguchi, G
影响因子：
64.8
作者：
STREISINGER, G;WALKER, C;SINGER, F
通讯作者：
SINGER, F