Establishment of germline-competent embryonic stem cell lines from the MSM/Ms strain
- 212 Downloads
MSM/Ms is an inbred mouse strain established from the Japanese wild mouse, Mus musculus molossinus, which has been phylogenetically distinct from common laboratory mouse strains for about 1 million years. The nucleotide substitution rate between MSM/Ms and C57BL/6 is estimated to be 0.96%. MSM/Ms mice display unique characteristics not observed in the commonly used laboratory strains, including an extremely low incidence of tumor development, high locomotor activity, and resistance to high-fat-diet-induced diabetes. Thus, functional genomic analyses using MSM/Ms should provide a powerful tool for the identification of novel phenotypes and gene functions. We report here the derivation of germline-competent embryonic stem (ES) cell lines from MSM/Ms blastocysts, allowing genetic manipulation of the M. m. molossinus genome. Fifteen blastocysts were cultured in ES cell medium and three ES lines, Mol/MSM-1, -2, and -3, were established. They were tested for germline competency by aggregation with ICR morulae and germline chimeras were obtained from all three lines. We also injected Mol/MSM-1 ES cells into blastocysts of ICR or C57BL/6 × BDF1 mice and found that blastocyst injection resulted in a higher production rate of chimeric mice than did aggregation. Furthermore, Mol/MSM-1 subclones electroporated with a gene trap vector were also highly efficient at producing germline chimeras using C57BL/6 × BDF1 blastocyst injection. This Mol/MSM-1 ES line should provide an excellent new tool allowing the genetic manipulation of the MSM/Ms genome.
KeywordsEmbryonic Stem Embryonic Stem Cell Bacterial Artificial Chromosome Embryonic Stem Cell Line Chimeric Mouse
This work was supported by KAKENHI (A) (17200028) and (B) (19300149) from the Japan Society for the Promotion of Science (JSPS). The MSM/Ms mouse strain (RBRC00209) was provided by RIKEN BRC, which is a participant in the National Bio-Resource Project of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. We thank Dr. M. Muta, Ms. Y. Tsuruta, and K. Haruna for their technical assistance, and the Center for Animal Resources and Development for the care of the animals used in this study.
- Araki K, Imaizumi T, Sekimoto T, Yoshinobu K, Yoshimuta J et al (1999) Exchangeable gene trap using the Cre/mutated lox system. Cell Mol Biol (Noisy-le-grand) 45:737–750Google Scholar
- Bonhomme F, Guénet JL (1996) The laboratory mouse and its wild relatives. In: Lyon MF, Rastan S, Brown SDM (eds) Genetic variants and strain of the laboratory mouse. Oxford University Press, Oxford, pp 1577–1596Google Scholar
- Lawitts JA, Biggers JD (1993) Culture of preimplantation embryos. In: Wasserman PM, Depamphilis ML (eds) Methods in enzymology, vol 225. Academic Press, San Diego, CA, pp 153–164Google Scholar
- Moriwaki K, Shiroishi T, Yonekawa H (1994) Genetics in wild mice. Japan Scientific Societies Press/Karger, Tokyo/BaselGoogle Scholar
- Nagy A, Gertsenstein M, Vintersten K, Behringer R (2003) Manipulating the mouse embryo. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
- Nakanishi M, Tazawa H, Tsuchiya N, Sugimura T, Tanaka T et al (2007) Mouse strain differences in inflammatory responses of colonic mucosa induced by dextran sulfate sodium cause differential susceptibility to PhIP-induced large bowel carcinogenesis. Cancer Sci 98:1157–1163CrossRefPubMedGoogle Scholar
- Robertson EJ (1987) Embryo-derived stem cell lines. In: Robertson EJ (ed) Teratocarcinomas and embryonic stem cells. A practical approach. IRL Press, Oxford, pp 71–112Google Scholar