EFFICIENT AND SUSTAINED TRANSGENE EXPRESSION IN MATURE RAT OLIGODENDROCYTES IN PRIMARY CULTURE

In order to evaluate the characteristics and efficiency of gene transf er in primary cultures of oligodendrocytes, four different techniques including particle bombardment (Accell(R) gene gun), cationic liposome -mediated transfection (lipofection), calcium phosphate co-precipitati on and retroviral infection were compared using the LacZ and luciferas e reporter genes, Highly purified postnatal adult rat oligodendrocytes were obtained by sequential immunopanning, plated in culture, and tra nsfected using various reporter and promoter genes, The most efficient expression of LacZ and luciferase genes was found with particle media ted gene delivery, The transgene expression level obtained with gene g un delivery was at least two- to 100-fold greater than three other tes ted gene transfer methods, Comparison of the relative strength of four viral and two cellular promoters in these primary oligodendrocytes cu ltures demonstrated that the CMV promoter was the strongest, Using a h uman growth hormone (hGH) reporter gene, a long-term transgene express ion pattern in primary oligodendrocytes was demonstrated to be sustain ed in culture for the entire experimental period (4 weeks) after parti cle-mediated gene transfer, These results demonstrate that expression of a foreign gene can be effectively achieved in primary cultures of a dult oligodendrocytes, especially by using the particle bombardment me thod, The results also suggest that the current ex vivo gene transfer system may be used to manipulate oligodendrocytes for future applicati on in gene therapy studies, (C) 1996 Wiley-Liss, Inc.