Derivation
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The R1/E cell line was subcloned from R1 in EMBL, Heidelberg, Germany by Kristina Vintersten. The R1 cell line was established in August 1991, from a 129X1 x 129S1 3.5 day blastocyst.
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Comments
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The R1/E cell line was subcloned from R1 in EMBL, Heidelberg, Germany by Kristina Vintersten. The R1 cell line was established in August 1991, from a 129X1 x 129S1 3.5 day blastocyst. The cells are heterozygous for the c locus (+/c (ch)) and for the pink eye locus (+/p).
This mouse ES cell line has been shown to be germline competent.In the F1 generation the coat color is uniform agouti, while in the F2 these two coat color genes segregate. The segregation could result in several coat types, from albino, through light brown, to black, depending on the genetic background of the partner of the germline chimaera.
Pluripotency of R1 was initially tested by tetraploid embryo <-> ES aggregates for completely ES derived development [PubMed: 8378314]. They were also tested by diploid embryo <-> ES aggregates and blastocyst injection for germline transmission in chimeras [PubMed: 8361547]. At early passages (up to passage #14), one third of the completely R1-derived newborns generated by tetraploid embryo <-> R1 aggregates survived. No live offspring were produced from cells older than passage #14.
However, about 20% of subclones derived from passage #14 had the original developmental potential of R1 when tested by tetraploid aggregates [PubMed: 8378314]. R1-derived animals reached adulthood and were fertile. The genetically altered lines derived from R1 gave high efficiency of germline transmission either by injecting them into C57 blastocyst or aggregating them with CD-1 or ICR outbred 8-cell stage embryos. More than 90% of the individual K.O. clones went to germline (n>60) by aggregation chimeras.
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