Efficient differentiation of embryonic stem cells into mesodermal precursors by BMP, retinoic acid and Notch signalling.

PloS One
Josema TorresFiona M Watt

Abstract

The ability to direct differentiation of mouse embryonic stem (ES) cells into specific lineages not only provides new insights into the pathways that regulate lineage selection but also has translational applications, for example in drug discovery. We set out to develop a method of differentiating ES cells into mesodermal cells at high efficiency without first having to induce embryoid body formation. ES cells were plated on a feeder layer of PA6 cells, which have membrane-associated stromal-derived inducing activity (SDIA), the molecular basis of which is currently unknown. Stimulation of ES/PA6 co-cultures with Bone Morphogenetic Protein 4 (BMP4) both favoured self-renewal of ES cells and induced differentiation into a Desmin and Nestin double positive cell population. Combined stimulation with BMP4 and all-trans Retinoic Acid (RA) inhibited self-renewal and resulted in 90% of cells expressing Desmin and Nestin. Quantitative reverse transcription-polymerase chain reaction (qPCR) analysis confirmed that the cells were of mesodermal origin and expressed markers of mesenchymal and smooth muscle cells. BMP4 activation of a MAD-homolog (Smad)-dependent reporter in undifferentiated ES cells was attenuated by co-stimulation with RA ...Continue Reading

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Citations

Dec 31, 2015·Journal of Muscle Research and Cell Motility·Barbara ŚwierczekKarolina Archacka
Dec 17, 2014·Stem Cells·Federico GalvagniSalvatore Oliviero
Apr 14, 2016·Expert Opinion on Biological Therapy·Kian Lam LimTunku Kamarul
Aug 17, 2018·Development·Amanda JanesickBruce Blumberg
Dec 23, 2015·Molecular Systems Biology·Hanna L Sladitschek, Pierre A Neveu
Jul 3, 2019·Stem Cells and Development·Kyle BurrellIna Dobrinski
Sep 27, 2014·The Journal of Biological Chemistry·Pooja AgrawalRobert E Hughes
Dec 31, 2019·Molecular Systems Biology·Hanna L Sladitschek, Pierre A Neveu

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Methods Mentioned

BETA
flow cytometry
transfection
confocal microscopy
PCR

Software Mentioned

ASW
FlowJo
Statistical Package for the Social Sciences ( SPSS )
Adobe Photoshop
Adobe Illustrator
Excel
FV10
TreeStar

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