Tsutsumi Hideki, Inoue Ryo, Yasuda Masahiko, Takahashi Riichi, Suzuki Masami, Urano Koji
Central Institute for Experimental Animals (CIEA), 3-25-12 Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa 210-0821, Japan.
J Toxicol Pathol. 2022 Jan;35(1):19-24. doi: 10.1293/tox.2021-0068. Epub 2021 Nov 19.
The rasH2 mouse was developed as a model for carcinogenicity studies in regulatory science. Its phenotype is stable during high-volume production and over successive generations. To produce rasH2 mice, three strains of mice (C57BL/6J-TgrasH2, C57BL/6J, and BALB/cByJ) were maintained individually. Since the homozygous genotype is lethal, hemizygous transgenic mice were maintained by crossing with inbred C57BL/6J mice. After breeding, male B6-transgenic mice were mated with female BALB/cByJ mice to obtain transgenic mice. Pups that were rasH2-Tg (tg/wt) or rasH2-Wt (wt/wt) were confirmed by genotyping. Frozen embryos were preserved by the Central Institute for Experimental Animals (CIEA) and sent to two facilities, CLEA Japan and Taconic Biosciences, where the mice were produced. Production colonies are created in both facilities and supplied to customers worldwide. To prevent genetic drift, the colonies were renewed for up to 10 generations, and renewals were carried out four times every five years from 2005 to 2021. To ensure the uniformity and maintenance of the phenotype of rasH2 mice, the carcinogen susceptibilities were monitored in every renewal of colonies by CIEA based on a standard protocol of the short-term carcinogenicity study using the positive control compound N-methyl-N-nitrosourea (MNU). Furthermore, simple carcinogenicity monitoring targeting the forestomach, the organ most sensitive to MNU, was performed approximately once a year. Based on the optimally designed production and monitoring systems, the quality of rasH2 mice with reproducibility and stability of carcinogenicity is maintained and supplied globally.
rasH2小鼠是作为监管科学中致癌性研究的模型而培育出来的。其表型在大规模生产过程中以及连续几代中都保持稳定。为了培育rasH2小鼠,三种小鼠品系(C57BL/6J-TgrasH2、C57BL/6J和BALB/cByJ)分别进行饲养。由于纯合基因型是致死性的,因此通过与近交系C57BL/6J小鼠杂交来维持半合子转基因小鼠。繁殖后,雄性B6转基因小鼠与雌性BALB/cByJ小鼠交配以获得转基因小鼠。通过基因分型确认rasH2-Tg(tg/wt)或rasH2-Wt(wt/wt)的幼崽。冷冻胚胎由实验动物中央研究所(CIEA)保存,并送往两个机构,即日本 CLEA 和 Taconic Biosciences,在那里培育小鼠。两个机构都建立了生产种群并供应给全球客户。为防止基因漂移,种群每10代更新一次,从2005年到2021年每五年进行四次更新。为确保rasH2小鼠表型的一致性和稳定性,CIEA根据使用阳性对照化合物N-甲基-N-亚硝基脲(MNU)的短期致癌性研究标准方案,在每次种群更新时监测致癌易感性。此外,大约每年对最易受MNU影响的器官——前胃进行一次简单的致癌性监测。基于优化设计的生产和监测系统,rasH2小鼠具有致癌性的可重复性和稳定性的质量得以维持并在全球范围内供应。