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

Aashir Awan; Roberto S Oliveri; Pernille L Jensen; Søren T Christensen; Claus Yding Andersen

doi:10.1007/978-1-60761-369-5_11

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

Aashir Awan, Roberto S Oliveri, Pernille L Jensen, Søren T Christensen, Claus Yding Andersen

Cellebiologi og Fysiologi

16 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	Methods in Molecular Biology
Vol/bind	584
Sider (fra-til)	195-210
Antal sider	16
ISSN	1064-3745
DOI	https://doi.org/10.1007/978-1-60761-369-5_11
Status	Udgivet - 1 jan. 2010

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10.1007/978-1-60761-369-5_11

Citationsformater

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

abstract = "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.",

keywords = "Base Sequence, Biological Markers, Cell Culture Techniques, Cell Differentiation, Cell Proliferation, Cilia, DNA Primers, Embryonic Stem Cells, Genetic Markers, Humans, Microscopy, Fluorescence, RNA, Messenger, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction",

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AU - Awan, Aashir

AU - Oliveri, Roberto S

AU - Jensen, Pernille L

AU - Christensen, Søren T

AU - Andersen, Claus Yding

PY - 2010/1/1

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N2 - 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.

AB - 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.

KW - Base Sequence

KW - Biological Markers

KW - Cell Culture Techniques

KW - Cell Differentiation

KW - Cell Proliferation

KW - Cilia

KW - DNA Primers

KW - Embryonic Stem Cells

KW - Genetic Markers

KW - Humans

KW - Microscopy, Fluorescence

KW - RNA, Messenger

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Signal Transduction

U2 - 10.1007/978-1-60761-369-5_11

DO - 10.1007/978-1-60761-369-5_11

M3 - Journal article

C2 - 19907979

SN - 1064-3745

VL - 584

SP - 195

EP - 210

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

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

Abstract

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