标题：Sysmex Partec 倍性分析仪三倍体斑马鱼的生产与代孕繁殖应用
Abstract:
We report for first time comparison of two approaches for zebrafish triploid production using cold shock and heat shock treatment. Subsequently, produced triploid zebrafish were used as a recipients for intraperitoneal transplantation of ovarian and testicular cells originating from vas:EGFP strain in order to verify their suitability for surrogate reproduction. Heat shock treatment was far more effective evaluated as success rate of triploid production and viability in comparison to cold shock. Triploids were produced with up to 100% efficiency in particular females. As expected, all triploids were males. Subsequently, germ cells transplantation revealed that triploids are suitable surrogate hosts. Production of donor-derived sperm was achieved in 23% and 16% of triploids transplanted by testicular and ovarian cells, respectively. Success of the transplantation was confirmed by positive GFP signal detected in gonads of dissected fish and stripped sperm. Germline transmission was confirmed by fertilization tests followed by PCR analysis of embryos. Reproductive success of germline chimera triploids evaluated as fertilization rate and progeny development was comparable to control groups.
Keywords: zebrafish, chromosome manipulation, triploid, germ cell transplantation, surrogate
reproduction
摘要：
我们首次报道了采用冷休克与热休克处理两种方法生产斑马鱼三倍体的对比研究。随后，将所获三倍体斑马鱼作为受体，用于腹腔移植来自vas:EGFP品系的卵巢与睾丸细胞，以验证其作为代孕载体的适用性。评估结果显示，热休克处理在三倍体生产成功率及存活率方面远优于冷休克处理。部分雌性个体的三倍体生产效率高达100%。不出所料，所有三倍体均为雄性。进一步的生殖细胞移植实验表明，三倍体斑马鱼适合作为代孕载体。
在睾丸和卵巢细胞移植的三倍体中，分别有23%和16%实现了供体精子的产生。通过在解剖的鱼和剥离的精子的性腺中检测到阳性GFP信号，证实了移植的成功。通过受精试验和胚胎PCR分析证实了种系传播。通过受精率和后代发育评估，种系嵌合体三倍体的繁殖成功率与对照组相当。
关键词：Sysmex Partec 倍性分析仪，斑马鱼、染色体操作、三倍体、生殖细胞移植、代孕
