RRC ID 2407
著者 Shinmen A, Honda A, Ohkawa M, Hirose M, Ogonuki N, Yuzuriha M, Miki H, Mochida K, Inoue K, Abe K, Ito M, Ogura A.
タイトル Efficient production of intersubspecific hybrid mice and embryonic stem cells by intracytoplasmic sperm injection.
ジャーナル Mol Reprod Dev
Abstract Recently, mice and embryonic stem (ES) cells with allelic polymorphisms have been used extensively in the field of genetics and developmental biology. In this study, we examined whether intersubspecific hybrid mice and ES cells with these genotypes can be efficiently produced by intracytoplasmic sperm injection (ICSI). Frozen-thawed spermatozoa from wild-derived strains, JF1 (Mus musculus molossinus), MSM (M. m. molossinus), HMI (M. m. castaneus), and SWN (M. m. spp.), were directly injected into mature oocytes from laboratory mice ([C57BL/6 x DBA2]F1; M. m. domesticus). The in vitro and in vivo developmental capacity of F1 embryos was not significantly different among the groups (P > 0.05), and term offspring were efficiently obtained in all groups (27%-34% of transferred embryos). However, the mean body and placental weights of the offspring differed significantly with genotype (P < 5 x 10(-10)), with the HMI hybrid greatest in both body and placental weights. In an application study using these F1 offspring, we analyzed their mitochondrial DNA using intersubspecific polymorphisms and found the consistent disappearance of sperm mitochondrial DNA in the F1 progeny. In a second series of experiments, we generated F1 blastocysts by injecting MSM spermatozoa into C57BL/6 oocytes and used them to generate hybrid ES cell lines. The ES cell lines were established at a high efficiency (9 lines from 20 blastocysts) and their allelic polymorphisms were confirmed. Thus, ICSI using cryopreserved spermatozoa allows the efficient and immediate production of a number of F1 hybrid mice and ES cell lines, which can be used for polymorphic analysis of mouse genetics.
巻・号 74(9)
ページ 1081-8
公開日 2007-9-1
DOI 10.1002/mrd.20612
PMID 17290420
MeSH Animals Cell Line* Chimera* Cryopreservation DNA, Mitochondrial / genetics Embryonic Development Embryonic Stem Cells* Embryonic Structures Female Genotype Male Mice Mice, Mutant Strains* / anatomy & histology Mice, Mutant Strains* / genetics Phenotype Polymorphism, Genetic Semen Preservation Sperm Injections, Intracytoplasmic* Spermatozoa / cytology Spermatozoa / physiology
IF 2.823
引用数 10
WOS 分野 REPRODUCTIVE BIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY DEVELOPMENTAL BIOLOGY CELL BIOLOGY
リソース情報
実験動物マウス RBRC00209 RBRC00639 RBRC00654 RBRC00657