CRYOPRESERVATION OF SEMEN AND SOMATIC CELLS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。来自已知或疑似人类遗传病动物模型的携带者或受影响的狗的精液被收集、稀释、冷冻和储存以备将来使用。体细胞，通常是成纤维细胞，在培养中生长，收获，并在液氮中冷冻，以便稍后重新培养和通过多种方法进行研究，包括酶分析和DNA Southern印迹/聚合酶链式反应(DNA Southern blotting/PCR)扩增，用于连锁研究和细胞遗传学研究。猫精液样本由华盛顿特区国家动物园史密森研究所基因组资源库的JoGayle Howard和Budda Pukazheathi博士收集、冷冻和储存。冷冻的猫胚胎是由辛辛那提动物园的威廉·斯旺森博士与他的NCRR赠款合作制造和储存的。我们还为拉尔夫·布林斯特博士和伊娜·多布林斯基博士提供了狗和猫的睾丸，用于跨物种生殖细胞移植研究。我们保存了密歇根州立大学玛格丽特·琼斯博士提供的患有β-甘露糖症和粘多糖病IIID的山羊的精液和成纤维细胞。这些克隆受到琼斯博士退休的威胁，精液和成纤维细胞资源正被储存在液氮中。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Semen from carriers or affected dogs affected with genetic diseases that are known or suspected animal models of human genetic disease is collected, diluted, frozen, and stored for the future use. Somatic cells, usually fibroblasts, are grown in culture, harvested, and frozen in liquid nitrogen for later re-culture and study by a number of methods including enzyme assays and DNA Southern blotting/pcr amplification of specified markers for linkage studies, as well as cytogenetic studies. Feline semen samples are collected, frozen, and stored by Drs. JoGayle Howard and Budda Pukazheathi, Genome Resources Bank, Smithsonian Institution, National Zoological Park, Washington DC. Frozen feline embryos are made and stored by Dr. William Swanson, Cincinnati Zoo, in collaboration with his NCRR grant. We have also provided testes from dogs and cats to Drs. Ralph Brinster and Ina Dobrinski for cross-species germ cell transplantation studies. We store semen and fibroblasts from goats with beta-mannosidosis and mucopolysaccharidosis IIID provided by Dr. Margaret Jones, Michigan State University. These colonies were threatened by Dr. Jones retirement, and the semen and fibroblast resources are being stored in liquid nitrogen.