A novel ES cell line, TT2, with high germline-differentiating potency.
In producing mutant mice by gene-targeting and gene-trapping in embryonic stem (ES) cells, the efficient colonization of the mutant ES cells into germline is still a critical matter. We have established a new line of ES cells, TT2, from an F1 embryo between a C57BL/6 female and a CBA male. When the TT2 cells were injected into blastocysts, the colonization into each tissue was very low. However, when injected into eight-cell embryos, the cells segregated inside the blastomeres, localized in an inner cell mass of blastocysts developed 1 day later, and colonized efficiently in each tissue of the pups. The pups were disproportionately male, about half of which were composed of TT2-derived cells primarily; in more than 70% of the males, TT2-derived cells were dominant, accounting for over half of the total cells. When these males were mated, they exclusively yielded TT2-derived offspring. The germline-differentiating potency was stable during 3 weeks of culture. Twenty-one of 24 mutant clones independently isolated yielded germline chimeras, and 19 clones yielded them in a rate comparable to that of the parent cells. Thus, TT2 cells can serve as a valuable vehicle for the production of mutant mice.
Yagi
T
T
Laboratory of Molecular Oncology, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.
Tokunaga
T
T
Furuta
Y
Y
Nada
S
S
Yoshida
M
M
Tsukada
T
T
Saga
Y
Y
Takeda
N
N
Ikawa
Y
Y
Aizawa
S
S
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D015139:Blotting, Southern; D002454:Cell Differentiation; D002460:Cell Line; D002678:Chimera; D002874:Chromosome Mapping; D046508:Culture Techniques / Q000379:methods; D004247:DNA / Q000032:analysis; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D005821:Genetic Techniques; D005854:Germ Cells / Q000166:cytology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008815:Mice, Inbred Strains; D008817:Mice, Mutant Strains; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D012727:Sex Characteristics; D012744:Sex Ratio; D013234:Stem Cells / Q000166:cytology*
D004247:DNA