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. 2011 Oct 13;21(9):1571–1586. doi: 10.1089/scd.2011.0370

FIG. 4.

FIG. 4.

Chimera formation after injection of rat 4.5 dpc blastocysts. (A) ESC line 3 cells were nucleofected with the pCX-eGFP plasmid and expanded to passage 11. These cells were injected into non-eGFP, parental strain F344 4.5 dpc blastocysts. Injected blastocysts were cultured for 10 days on inactivated mouse embryonic fibroblast feeder (MEF) cells after injection (top) or after 14 days (bottom) of culture on inactivated MEFs. The same field is shown in each panel using phase-contrast illumination (left), GFP fluorescence (middle), and a merged image is shown on the right. Scale Bar 100 μm. (B) A single female chimera from a litter of 9 pups after injection of the male line 54.1 into F344 blastocysts. (C) Left: 4 chimera from a litter of 5 pups after injection of male line 53.1 into SD blastocysts. Note that this ESC line demonstrated germline transmission. Note that there is a variation in the degree of chimerism for each ESC line. Middle and right: 4 chimera from 2 litters of 3 pups after injection of male line 52 into SD blastocysts. (D) PCR microsatellite genotyping data after injection of male F344 line into SD blastocysts (lanes 1–4 on left and lines 1–4 on right). Lanes 5–8 are control samples for F344, DA, DA× SD, and SD, respectively. Note that 3 F344× SD chimera are indicated by astrisks in gel on the right. eGFP, enhanced green fluorescent protein; dpc, days post coitum; SD, Sprague-Dawley.