Neural differentiation of pluripotent mouse embryonal carcinoma cells by retinoic acid: inhibitory effect of serum
Language English Country Czech Republic Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Antineoplastic Agents pharmacology MeSH
- Biomarkers MeSH
- Cell Differentiation drug effects physiology MeSH
- Gene Expression drug effects MeSH
- Blood Proteins pharmacology MeSH
- Culture Media, Serum-Free pharmacology MeSH
- Cells, Cultured MeSH
- Mice MeSH
- Neurons cytology MeSH
- Pluripotent Stem Cells cytology drug effects physiology MeSH
- Response Elements physiology MeSH
- Tretinoin pharmacology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Antineoplastic Agents MeSH
- Biomarkers MeSH
- Blood Proteins MeSH
- Culture Media, Serum-Free MeSH
- Tretinoin MeSH
In both embryonal carcinoma (EC) and embryonic stem (ES) cells, the differentiation pathway entered after treatment with retinoic acid (RA) varies as it is based upon different conditions of culture. This study employs mouse EC cells P19 to investigate the effects of serum on RA-induced neural differentiation occurring in a simplified monolayer culture. Cell morphology and expression of lineage-specific molecular markers document that, while non-neural cell types arise after treatment with RA under serum-containing conditions, in chemically defined serum-free media RA induces massive neural differentiation in concentrations of 10(-9) M and higher. Moreover, not only neural (Mash-1) and neuroectodermal (Pax-6), but also endodermal (GATA-4, alpha-fetoprotein) genes are expressed at early stages of differentiation driven by RA under serum-free conditions. Furthermore, as determined by the luciferase reporter assay, the presence or absence of the serum does not affect the activity of the retinoic acid response element (RARE). Thus, mouse EC cells are able to produce neural cells upon exposure to RA even without culture in three-dimensional embryoid bodies (EBs). However, in contrast to standard EBs-involving protocol(s), neural differentiation in monolayer only takes place when complex signaling from serum factors is avoided. This simple and efficient strategy is proposed to serve as a basis for neurodifferentiation studies in vitro.
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