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 language | English (US) |
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Journal | Current Protocols in Human Genetics |
Issue number | SUPPL. 56 |
DOIs | |
State | Published - 2008 |
Keywords
- Hematopoietic
- Progenitors
- Retroviral vector
- Transduction
ASJC Scopus subject areas
- Genetics
- Genetics(clinical)