Procedures for Derivation and Characterisation of Human Embryonic Stem Cells from Odense, Denmark

Linda Harkness, Moustapha Kassem

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearch

Abstract

In 1998, a development occurred in stem cell biology with the fi rst report of the derivation of a human embryonic stem cell (hESC) line. Since then a number of techniques have been used to derive and characterise hESCs. Here, we describe the derivation methods used by our laboratory for isolation of the ICM by immunosurgery and outgrowth of the whole blastocyst. We have added protocols for routine culture, passaging and cryopreservation of our hESC lines as well as the methods we have used for characterisation (flow cytometry, karyotyping, immunocytochemistry, in vitro and in vivo differentiation). Additionally, we have included gene sequences for PCR and an antibody list for immunocytochemistry.
Original languageEnglish
Title of host publicationHuman Embryonic Stem Cells Handbook : Methods in Molecular Biology
EditorsKursad Turksen
PublisherSpringer Science+Business Media
Publication date2012
Pages33-51
Chapter3
ISBN (Print)978-1-61779-794-1
DOIs
Publication statusPublished - 2012
SeriesMethods in Molecular Medicine
Volume873
ISSN1543-1894

Keywords

  • Human embryonic stem cells
  • Derivation
  • Characterisation

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