Developmental potential of teratocarcinoma stem cells in utero following aggregation with cleavage-stage mouse embryos

Embryonal carcinoma cells were aggregated with cleavage stage mouse embryos, cultured briefly, and transferred as morulae to the uteri of pseudopregnant females. When midgestation foetuses were examined, many were morphologically abnormal. The severity of this abnormal development was correlated with the extent of contribution by embryonal carcinoma cells to the foetuses as indicated by GPI (glucose phosphate isomerase) analysis. This was true for all three of the cell lines studied, NG-2, PSA-1, and LT1-2D. The clear correlation between increasingly abnormal development and more extensive participation by the embryonal carcinoma cells was not observed in control experiments in which embryos of different stages of development were aggregated together. The data therefore suggest that the embryonal carcinoma cells studied here are unable to support normal development in the absence of a substantial number of host embryonic cells. It remains unclear whether this is a consequence of the karyotypic abnormalities of the cells tested, or whether it reflects a characteristic limitation in the ability of embryonal carcinoma cells to independently direct normal development. When aggregates were allowed to develop to term and the extent of chimaerism was examined in the live-born animals, it was found to be sporadic and limited. This is consistent with the results indicating that large contributions by embryonal carcinoma cells are not compatible with normal development at midgestation. The chimaerism observed in the live-born animals was comparable in both frequency and in tissue distribution to that generally obtained in other studies using either the aggregation or blastocyst injection techniques.