Cell mixing during the early development of mouse aggregation chimera
Published: 1 August 1995
P Dvorak, A Yoshiki, D Dvorakova, J E Flechon and M Kusakabe
Laboratoire de Biologie Cellulaire et Moleculaire, INRA, Jouy en Josas, France.
Two different inbred strain combinations of mouse aggregation chimeras - C3H/HeN (H-2k) x C57BL/6N (H-2b) and C3H/HeN x BALB/cA (H-2d) were used for cell mixing analysis at two points in time - 24 h after aggregation (just prior to transplantation into foster mothers) and 7.5 days post coitum (p.c.). The cell proportion of two H-2 haplotypes at the blastocyst stage was studied using a fluorescence-labeled monoclonal antibody recognizing a C3H-specific alloantigen - CSA (C3H strain-specific antigen) and laser scanning confocal microscopy. The 7.5-day-old chimeras were sectioned and subsequently processed by sensitive biotinylated antibody - avidin peroxidase immunohistochemical technique. Our results showed that 24 h after aggregation (blastocyst stage), there was equal cell mixing and no mouse strain used in the present study was dominant at this time. In 7.5-day-old C3H/HeN x BALB/cA chimeras, cells of both genotypes were intermingled, but the C3H/HeN strain was dominant in all cases. In contrast, the combination C3H/HeN x C57BL/6N clearly showed reduced numbers of C3H/HeN cells (CSA-positive) in 83% of the chimeras evaluated. Generally, CSA positive cells were found only in randomly distributed small distinct areas representing less than 20% of embryonal cells. Surprisingly, the extraembryonal ectoplacental cone was uniformly CSA positive in some C3H/HeN x C57BL/6N chimeras. Furthermore, in 36% of normally implanted chimeras of both strain combinations progressive degeneration was observed. We suggest that the cell mixing pattern as well as the absolute number of cells derived from each strain in the aggregation chimera can be affected by specific immune interactions involving H-2 haplotype combinations of the allogeneic fetus and the fully immunocompetent host organism, at later points in development.