Global microRNA analysis in primary hepatocyte cultures

Julian Krauskopf, Almudena Espín-Pérez, Jos C. Kleinjans, Theo M. De Kok

Research output: Chapter in Book/Report/Conference proceedingChapterAcademic

Abstract

MicroRNAs are small non-coding molecules that regulate gene expression and in return affect diverse biological functions, including those involved in toxicity and development of disease. Recent evidence suggests that microRNAs play an important role in liver pathologies, like viral hepatitis, alcoholic liver, hepatocellular carcinoma, or drug-induced liver injury. Furthermore, numerous studies demonstrated the high potential of microRNAs as promising non-invasive biomarkers of liver disease or as relevant targets for therapeutic treatment. This chapter describes a method for global microRNA analysis of primary hepatocytes by high-throughput sequencing. The method comprises the isolation of high-quality total RNA, analysis of microRNA sequencing data, and the validation of the findings by reverse transcriptase quantitative polymerase chain reaction analysis.
Original languageEnglish
Title of host publicationProtocols in In Vitro Hepatocyte Research
PublisherSpringer Verlag
Pages241-250
Number of pages10
ISBN (Print)9781493920747
DOIs
Publication statusPublished - 23 Jan 2015

Publication series

SeriesProtocols in In Vitro Hepatocyte Research

Keywords

  • MicroRNAs
  • Primary hepatocyte
  • Small RNA sequencing
  • Total RNA isolation

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