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Region Hovedstaden - en del af Københavns Universitetshospital
Udgivet

Immunoflourescence and mRNA analysis of human embryonic stem cells (hESCs) grown under feeder-free conditions

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  1. Determination of Binding Kinetics of Intrinsically Disordered Proteins by Surface Plasmon Resonance

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  2. Analysis of Mass Cytometry Data

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  3. Assessment of Peptidylarginine Deiminase Activity by ELISA Using Human Fibrinogen as Substrate

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  4. Full-Length Open Reading Frame Amplification of Hepatitis C Virus

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  5. In Vitro Neutralization Assay Using Cultured Hepatitis C Virus

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  1. Expression of the Insulin-like Growth Factor System in First- and Second-Trimester Human Embryonic and Fetal Gonads

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  2. Shortcomings of an unphysiological triggering of oocyte maturation using human chorionic gonadotropin

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  3. Surrogate testes: Allogeneic spermatogonial stem cell transplantation within an encapsulation device may restore male fertility

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  4. Effect of sphingosine-1-phosphate on activation of dormant follicles in murine and human ovarian tissue

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This chapter describes the procedures in order to do immunofluorescence (IF) microscopy and quantitative PCR (qPCR) analysis of human embryonic stem cells (hESCs) grown specifically under feeder-free conditions. A detailed protocol outlining the steps from initially growing the cells, passaging onto 16-well glass chambers, and continuing with the general IF and qPCR steps will be provided. The techniques will be illustrated with new results on cellular localization of transcriptional factors and components of the Hedgehog, Wnt, and PDGF signaling pathways to primary cilia in stem cell maintenance and differentiation. Furthermore, a sample qPCR experiment will be shown illustrating that these techniques can be important tools in answering basic questions about hESC biology.
OriginalsprogEngelsk
TidsskriftMethods in molecular biology
Vol/bind584
Sider (fra-til)195-210
Antal sider16
DOI
StatusUdgivet - 1 jan. 2010

ID: 31026799