Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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.
Original language | English |
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Title of host publication | Human Embryonic Stem Cell Protocols |
Volume | 1307 |
Publisher | Springer Science+Business Media |
Publication date | 2016 |
Pages | 123-140 |
ISBN (Print) | 978-1-4939-2667-1, 978-1-4939-2668-8 |
DOIs | |
Publication status | Published - 2016 |
Series | Methods in molecular biology (Clifton, N.J.) |
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ISSN | 1064-3745 |
ID: 162378679