Expression of transforming growth factor beta 2 during the differentiation of murine embryonal carcinoma and embryonic stem cells.
Transforming growth factor beta 2 (TGF beta 2) mRNA expression was studied by Northern blot analysis in a range of feeder-independent murine embryonal carcinoma (EC) cells and in feeder-dependent EC and embryonic stem (ES) cells. TGF beta 2 transcripts were not detected in any undifferentiated cells including P19, F9, PC13, C1003, PSA-1, P10, and ES. Following induction of differentiation, however, TGF beta 2 became expressed, independently of the cell type formed. Retinoic acid (RA) addition and/or deprivation of the differentiation inhibiting activity of feeder cells resulted in the appearance of TGF beta 2 transcripts within 2 days. These kinetics correlated entirely with the first appearance of the protein; an anti-peptide antibody specifically recognizing TGF beta 2 did not stain P19 EC cells by immunofluorescence but 2-3 days after RA addition, a significant proportion of the population was strongly labeled. In addition, primitive endoderm cells emerging from the inner cell mass of substrate attached blastocysts stained brightly with anti-TGF beta 2, while the undifferentiated inner cell mass cells did not. Although all trophectoderm cells at the mid-blastocyst stage were stained, few had detectable levels of TGF beta 2 after plating on a substrate. Neither TGF beta 1 nor TGF beta 2 affected the growth of EC cells, but a range of differentiated derivatives were all inhibited, with TGF beta 2 being marginally more effective than TGF beta 1 at the same concentration.
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht.
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Mummery
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Slager
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Kruijer
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Feijen
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Freund
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Koornneef
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van den Eijnden-van Raaij
D000818:Animals; D015152:Blotting, Northern; D002454:Cell Differentiation; D002460:Cell Line; D004622:Embryo, Mammalian / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005796:Genes; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics*; D013724:Teratoma; D014158:Transcription, Genetic; D015291:Transforming Growth Factors / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism
D012333:RNA, Messenger; D015291:Transforming Growth Factors