[Gene-transfer into bone marrow cells].
We have intended to improve gene-transfer technique into hematopoietic stem cells for somatic gene therapy. 1) We have developed a new packaging cell line, ampGPE for retroviral production. LTR-less gag, pol or env genes from Moloney murine leukemia virus were separately inserted into BMGNeo vector. Packaging cell lines containing 20-50 copies of these two kinds of plasmid were obtained. Retrovirus stock for gene-transfer have been produced at a high titer (10(5)-10(6) cfu/ml) and without replication-competent viruses by using ampGPE. 2) Retrovirus-transduced murine CFU-GM have been found to selectively proliferate (5-10 fold/week) in liquid capture with recombinant murine IL-3, human IL-6 and G418 to consequently obtain enough amount of, highly concentrated (70-100%), and gene-transferred murine CFU-GM for gene-delivery system.
Takahara
Y
Y
Central Research Laboratories, Ajinomoto Co. Inc., Kawasaki, Japan.
Hamada
K
K
Okano
A
A
Takagi
T
T
Shimotono
K
K
D000818:Animals; D002460:Cell Line; D015316:Genetic Therapy; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D006412:Hematopoietic Stem Cells; D006801:Humans; D007377:Interleukin-3; D015850:Interleukin-6; D009355:Neomycin; D010957:Plasmids; D012190:Retroviridae / Q000235:genetics; D014162:Transfection
D007377:Interleukin-3; D015850:Interleukin-6; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D009355:Neomycin