Genetic modification of human hematopoietic cells: preclinical optimization of oncoretroviral-mediated gene transfer for clinical trials.
This chapter provides information about the oncoretroviral transduction of human hematopoietic stem/ progenitor cells under clinically applicable conditions. We describe in detail a short -60 h transduction protocol which consistently yields transduction efficiencies in the range of 30-50% with five different oncoretroviral vectors. We discuss a number of parameters that affect transduction efficiency, including the oncoretroviral vector characteristics, the vector stock collection, the source of CD34+ cells and transduction conditions.
Budak-Alpdogan
Tulin
T
Department of Medicine, University of Medicine and Dentistry of New Jersey, New Brunswick, NJ, USA.
Rivière
Isabelle
I
D018952:Antigens, CD34 / Q000276:immunology; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D016133:Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic
D018952:Antigens, CD34; D016895:Culture Media, Serum-Free