Cell surface markers to monitor the process of visceral endoderm differentiation from embryonal carcinoma cells: identification of the stage sensitive to high concentration of retinoic acid

Two cell surface antigens, brushin and FT-1 were effective in analysis of the process of visceral endoderm differentiation. Brushin was detected on both primitive and visceral endoderm, while FT-1 was detected only on visceral endoderm. When aggregates of N4-1 embryonal carcinoma cells were exposed to 10(-8) M-retinoic acid for more than 2 days, external cells differentiated to multilayered and vacuolized visceral endoderm. However, aggregates treated with 10(-6) M-retinoic acid developed an endoderm layer, which remained one cell thick and was not vacuolized. Cell surface properties of the endoderm cells indicated that the high concentration of retinoic acid inhibited the differentiation pathway at the stage between primitive endoderm cells and visceral endoderm cells. By pulsed exposure to 10(-6) M-retinoic acid, the period sensitive to the high concentration of retinoic acid was shown to be around day 4 after the initial exposure to retinoic acid.