Liposome-Mediated Gene Transfer into Established CNS Cell Lines, Primary Glial Cells, and in Vivo

Cell Transplantation

Original Contribution

Günter Stockhammer¹  Gottfried Baier²  Christine Rehrl²  Bettina Wiesenhofer³  Christian Humpel³  Patrick Kofler³ 

[1] Department Neurology, University Hospital Innsbruck, Innsbruck, Austria;Institute of Medical Biology and Human Genetics, University Innsbruck;Neurochemical Unit, Department Psychiatry, University Hospital Innsbruck;

关键词: Gene transfer;    Lipofection;    Reportergene;    Gene therapy;   

DOI  :  10.1177/096368979800700212

来源: Sage Journals

【 摘 要 】

Sufficient gene transfer into CNS-derived cells is the most crucial step to develop strategies for gene therapy. In this study liposome-mediated gene transfer using a β-galactosidase (β-GAL) reporter gene was performed in vitro (C6 glioma cells, NT2 neuronal precursor cells, 3T3 fibroblasts, primary glial cells) and in vivo. Using Trypan blue exclusion staining, optimal lipid concentration was observed in the range of 10-12 μg/mL. Under optimal conditions (80,000 cells/16 mm well, incubation overnight, lipid/DNA ratio = 1:18) a high transfection rate was achieved (<9% for C6 cells; <1% for NT2 cells). In primary cultures of glial cells a fair amount of positive stained cells (glial cell) was found, but the transfection efficiency was lower (<0.1%). A “boost-lipofection” markedly increased (twice) lipofection efficiency in C6 cells. Expression of β-GAL reached a maximum after 3-5 days. When the liposome–DNA complexes were injected/infused directly into the brains of adult rats, several weakly stained cells could be observed in the brain region adjacent to the injection site. It is concluded that liposome-mediated gene transfer is an efficient method for gene transfer into CNS cells in vitro, but the transfection efficiency into the rat brain in vivo is far too low and therefore not applicable.

【 授权许可】

Unknown   
© 1998 Elsevier Science Inc.