Implantation of BDNF-producing packaging cells into brain.
In order to invent a screening system to check in vivo gene function and the efficiency of gene transfer mediated by a retroviral vector system, we established a novel packaging cell, PacC6/A8, that is transplantable to rat brains. The packaging cell is based on the gene of the neuropatogenic retrovirus, A8-V. For expression in the brain, a vector that expresses brain-derived neurotrophic factor (BDNF) tagged by c-Myc-His6 (LxA/bdmh) was constructed. After transfection of LxA/bdmh to PacC6/A8, a cloned cell line, PacC6/A8/bmh, was established. PacC6/A8/bmh cells stably produced pseudotyped retroviruses carrying LxA/bdmh. For a control, a retroviral vector that bears the gene that codes enhanced green fluorescent protein (EGFP) tagged by C-Mic-His6 was also created and used for the establishment of PacC6/A8/gfmh cells that produce pseudotyped retroviruses carrying LxA/gfmh. PacC6/A8/bmh and PacC6/A8/gfmh cells were injected to the brain of newborn rats. A tumor was formed in all the rats injected that did not exhibit any symptoms until 3-4 weeks after the injection. A histological study of the injected rats revealed that the transferred BDNF gene was expressed in the brain of rats injected with PacC6/A8/bmh cells, but not in rats with PacC6/A8/gfmh cells. Interestingly, many activated microglia had migrated into the tumor induced by PacC6/A8/bmh cells, and expressed a high amount of BDNF.
Fukumitsu
Hidefumi
H
Institute of Life Science, Soka University, Hachioji, Tokyo, Japan.
Takase-Yoden
Sayaka
S
Furukawa
Shoei
S
Nemoto
Kiyomitsu
K
Ikeda
Tomio
T
Watanabe
Rihito
R
D000818:Animals; D001921:Brain / Q000033:anatomy & histology / Q000601:surgery*; D019208:Brain-Derived Neurotrophic Factor / Q000096:biosynthesis / Q000235:genetics*; D019556:COS Cells; D017690:Cell Transplantation / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins; D051381:Rats; D012190:Retroviridae / Q000235:genetics* / Q000302:isolation & purification; D014161:Transduction, Genetic / Q000379:methods; D014407:Tumor Cells, Cultured; D019065:Virus Assembly
D019208:Brain-Derived Neurotrophic Factor; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins