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. In 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 alpha-fetoprotein. In the present study, cellular autofluorescence profiles obtained by fluorescence-activated 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 flow cytometry provides a novel mechanism for the separation of undifferentiated cells from differentiated endoderm cells in F9 embryoid bodies.
Burdsal
C A
CA
Laboratory of Radiobiology and Environmental Health, University of California, San Francisco, USA.
Pedersen
R A
RA
Hyun
W C
WC
Latimer
J J
JJ
D000818:Animals; D000906:Antibodies; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D005456:Fluorescent Dyes; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D012313:RNA / Q000032:analysis; D012334:RNA, Neoplasm / Q000032:analysis; D011817:Rabbits / Q000276:immunology; D018243:Teratocarcinoma; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014909:Wheat Germ Agglutinins
D000906:Antibodies; D005456:Fluorescent Dyes; D012334:RNA, Neoplasm; D014909:Wheat Germ Agglutinins; C047939:fluorescein isothiocyanate-wheat germ agglutinin; D014212:Tretinoin; D012313:RNA; D016650:Fluorescein-5-isothiocyanate