Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways

Maj Linea Vestergaard; Aashir Awan; Caroline Becker Warzecha; Søren Tvorup Christensen; Claus Yding Andersen

doi:10.1007/7651_2014_127

Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways

Maj Linea Vestergaard, Aashir Awan, Caroline Becker Warzecha, Søren Tvorup Christensen, Claus Yding Andersen

11 Citationer (Scopus)

Abstract

This chapter describes the procedures for immunofluorescence microscopy (IFM) and quantitative PCR (qPCR) analyses of human embryonic stem cells (hESCs) grown specifically under feeder-free conditions. A detailed protocol is provided outlining the steps from initially growing the cells, passaging onto 16-well glass chambers, and continuing with the general IFM and qPCR anlysis. The techniques are illustrated with results on cellular localization of transcriptional factors and components of the Hedgehog, Wnt, PDGF, and TGFβ signaling pathways to primary cilia in stem cell maintenance and differentiation. Furthermore, a sample qPCR experiment is experimentally shown illustrating that these techniques can be important tools in answering basic questions about hESC biology.

Originalsprog	Engelsk
Titel	Human Embryonic Stem Cell Protocols
Vol/bind	1307
Forlag	Springer Science+Business Media
Publikationsdato	2015
Sider	123-140
ISBN (Trykt)	978-1-4939-2667-1, 978-1-4939-2668-8
DOI	https://doi.org/10.1007/7651_2014_127
Status	Udgivet - 2015

Navn	Methods in molecular biology (Clifton, N.J.)
ISSN	1064-3745

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10.1007/7651_2014_127

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Vestergaard, M. L., Awan, A., Warzecha, C. B., Christensen, S. T., & Andersen, C. Y. (2015). Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways. I Human Embryonic Stem Cell Protocols (Bind 1307, s. 123-140). Springer Science+Business Media. Methods in molecular biology (Clifton, N.J.) https://doi.org/10.1007/7651_2014_127

Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways. / Vestergaard, Maj Linea; Awan, Aashir; Warzecha, Caroline Becker et al.

Human Embryonic Stem Cell Protocols. Bind 1307 Springer Science+Business Media, 2015. s. 123-140 (Methods in molecular biology (Clifton, N.J.)).

Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › peer review

Vestergaard, ML, Awan, A, Warzecha, CB, Christensen, ST & Andersen, CY 2015, Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways. i Human Embryonic Stem Cell Protocols. bind 1307, Springer Science+Business Media, Methods in molecular biology (Clifton, N.J.), s. 123-140. https://doi.org/10.1007/7651_2014_127

Vestergaard ML, Awan A, Warzecha CB, Christensen ST , Andersen CY. Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways. I Human Embryonic Stem Cell Protocols. Bind 1307. Springer Science+Business Media. 2015. s. 123-140. (Methods in molecular biology (Clifton, N.J.)). doi: 10.1007/7651_2014_127

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Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways

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