Human hematopoietic cell culture, transduction, and analyses

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


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 for maintenance 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.

TidsskriftCurrent protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.]
Vol/bindChapter 13
Sider (fra-til)Unit 13.7
StatusUdgivet - jan. 2008


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