Human hematopoietic cell culture, transduction, and analyses

Jesper Bonde, Louisa Wirthlin, Donald B. Kohn, Jan Nolta

Research output: Contribution to journalArticle

Abstract

This unit provides methods for introducing genes into human hematopoietic progenitor cells. The Basic Protocol describes isolation of CD34+ cells, transduction of these cells with a retroviral vector on fibronectin-coated plates, assaying the efficiency of transduction, and establishing long-term cultures. Support protocols describe methods formaintenance of vector-producing fibroblasts (VPF) and supernatant collection from these cells, screening medium components for the ability to support hematopoietic cell growth, and establishing colonies from long-term cultures. Other protocols provide PCR-based methods to analyze individual colonies for transduction, methods to analyze cells harvested from long-term cultures, and procedures for freezing and thawing of hematopoietic cells.

Original languageEnglish (US)
JournalCurrent Protocols in Human Genetics
Issue numberSUPPL. 56
DOIs
StatePublished - 2008

Keywords

  • Hematopoietic
  • Progenitors
  • Retroviral vector
  • Transduction

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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