Removal of inhibitory substances with recombinant fibronectin-CH-296 plates enhances the retroviral transduction efficiency of CD34(+)CD38(-) bone marrow cells.
In retroviral gene transduction, the efficiency of viral infection was reduced by the proteoglycans and some other materials secreted by the producer lines. In order to remove these inhibitors we have developed the rFN-CH-296-facilitated protocol. Because the rFN-CH-296 molecule has strong ability to bind a retroviral vector, rFN-CH-296 bound plates are utilized to wash out the unbound putative inhibitors present in a virus supernatant. The gene transduction efficiencies of human CD34(+)CD38(-) BMCs with a GALV-pseudotyped vector and the rFN-CH-296-facilitated protocol were compared with the protocol without a coating plate with CH-296, the mean gene transduction efficiencies being found to be 95.5 and 71.1%, respectively.
Chono
H
H
Biotechnology Research Laboratories, Takara Shuzo Co., Ltd., Seta, Otsu, Shiga 520-2193, Japan.
Yoshioka
H
H
Ueno
M
M
Kato
I
I
D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000818:Animals; D015703:Antigens, CD; D018952:Antigens, CD34 / Q000378:metabolism; D000943:Antigens, Differentiation / Q000378:metabolism; D001665:Binding Sites / Q000502:physiology; D001854:Bone Marrow Cells / Q000378:metabolism* / Q000821:virology; D005353:Fibronectins / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics*; D008562:Membrane Glycoproteins; D051379:Mice; D009244:NAD+ Nucleosidase / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012192:Retroviridae Infections / Q000378:metabolism; D014161:Transduction, Genetic
D015703:Antigens, CD; D018952:Antigens, CD34; D000943:Antigens, Differentiation; D005353:Fibronectins; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011509:Proteoglycans; D011994:Recombinant Proteins; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; C498155:Cd38 protein, mouse; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1