Open AccessNovel flow‐cytometric method for separating cell types in differentiated F9 embryoid bodies
Author(s) -
Burdsal Carol A.,
Pedersen Roger A.,
Hyun William C.,
Latimer Jean J.
Publication year - 1995
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990210206
Subject(s) - embryoid body , endoderm , retinoic acid , biology , population , teratocarcinoma , microbiology and biotechnology , flow cytometry , p19 cell , embryonic stem cell , cellular differentiation , cell type , cell , cell culture , biochemistry , induced pluripotent stem cell , genetics , demography , sociology , gene
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 α‐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 α‐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. © 1995 Wiley‐Liss, Inc.