NOVEL FLOW-CYTOMETRIC METHOD FOR SEPARATING CELL-TYPES IN DIFFERENTIATED F9 EMBRYOID BODIES

The differentiation of F9 teratocarcinoma cells mimics the formation of a mouse embryonic tissue, the primitive endoderm, Tn vitro, small aggregates of F9 cells, termed embryoid bodies, differentiate in response to retinoic acid and develop a surface epithelium that is characterized by the production of cw-fetoprotein. In the present study, cellular autofluorescence profiles obtained by fluorescence-activated cell sorting demonstrated that undifferentiated embryoid bodies were composed of a single type of cell. In contrast, retinoic acid-induced embryoid bodies were composed of two cell types: a major population displaying autofluorescence levels similar to those of cells from undifferentiated embryoid bodies and a second population displaying higher autofluorescence. RNA analyses demonstrated that the transcription of alpha-fetoprotein was associated only with the more highly autofluorescent population, indicating that how cytometry provides a novel mechanism for the separation of undifferentiated cells from differentiated endoderm cells in F9 embryoid bodies. (C) 1995 Wiley-Liss, Inc.