Efficient preparation of cationized gelatin for gene transduction.

ABSTRACT

We previously reported gene therapy using cationized gelatin microspheres of φ20-32 µm, prepared from pig skin, as a transducing agent, but although the gelatin offered various advantages, its yield was extremely low (only 0.1%). In this study, we markedly improved the yield of φ20-32 µm cationized gelatin microspheres and prepared a newly less than φ20 µm cationized gelatin. Conventionally, cationized gelatin is prepared by cationization, particulation by agitation, and cross-linking. The yield is determined by the particulation step, for which we had used a three-necked distillation flask of 500 mL and an agitation speed of 420 rpm. The yield was significantly increased from 0.13 ± 0.02% to 8.80 ± 1.90% by using a smaller flask of 300 mL and an agitation speed of 25000 rpm (p ＜ 0.01). We could also prepare cationized gelatin of less than φ20 µm, which had not been possible previously. We confirmed that efficient gene introduction into peritoneal macrophages could be achieved with the new cationized gelatin.