Developing New Inbred Zebrafish Lines to Enhance Cell Transplantation Models

描述（由申请人提供）：本提案的长期目标是促进新型人类细胞疗法的实施。这种治疗方法的成功很大程度上取决于是否有可靠的动物模型来进行原理验证实验和临床前研究。斑马鱼已经在骨髓和皮肤等移植实验中进行了测试，然而，缺乏免疫兼容的鱼类严重阻碍了它的广泛应用。我们建议开发一种新的方法来产生移植相容的斑马鱼细胞系。这种新方法应该能够以低成本快速部署；潜在地利用广泛和快速扩展的突变体库来模拟人类疾病。我们的具体目标是：目标1，为生物医学社区使用的三种鱼类（即Tubingen， AB和Casper系）开发MHC兼容的斑马鱼。我们将通过体细胞核移植来实现这一目标。我们将从一条鱼身上克隆出雌雄同体，然后进行杂交；目的2，通过功能性研究证明这些动物在不使用免疫抑制的情况下耐受来自匹配供体的移植细胞的能力；我们将通过相互的皮肤移植来测试每一代动物的免疫相容性。我们正在解决该领域的一个关键障碍

DESCRIPTION (provided by applicant): The long-term goal of this proposal is to facilitate the implementation of novel human cell therapies. The success of such treatments much depends on having reliable animal models to perform proof of principle experiments and preclinical studies. Zebrafish have been tested in transplantation experiments such as bone marrow and skin, however, the lack of immune-compatible fish seriously hinders its broad use. We propose to develop a new approach to generate transplantation-compatible zebrafish lines. This new method should be capable of deploying rapidly, at low cost; potentially capitalizing on the extensive and rapidly-expanding library of mutants modeling human disease. Our specific aims are: Aim 1, to develop MHC compatible zebrafish for three strains of fish used by the biomedical community i.e. Tubingen, AB and Casper lines. We will accomplish this aim using somatic cell nuclear transfer. We will produce clones of both sexes from an individual fish that will be subsequently crossed; Aim 2, to demonstrate, using functional studies, the capacity of these animals to tolerate transplanted cells from matched donors without the use of immunosuppression; we will test animals of each generation for their immune compatibility using reciprocal skin transplants. We are addressing a critical barrier in the field by solving the problem of reliably generating immune- compatible zebrafish. In addition, the same strategy can be used for the generation of homozygous zebrafish lines in a considerably shorter period of time. New and already developed mutant zebrafish lines could be re-derived on a homozygous background enhancing their value as model organism.