Preparation of silk protein sericin as mitogenic factor for better mammalian cell culture

We previously reported that sericin small (sericin-S), with a molecular weight that ranges from 5 to 100 kDa, is a cell culture supplement used to accelerate cell proliferation. In this study, a novel preparation method for sericin and several applications of sericin were examined. Sericin large, prepared under nonhydrolyzing conditions and ranging from 50 to 200 kDa, also accelerated cell proliferation, but its effects were inferior to those of sericin-S. Additional sericin preparations with various molecular weights that were differentially hydrolyzed were also tested but none of them was significantly superior to sericin-S, and neither were several recombinant sericin peptides. Sericin-S successfully accelerated the proliferation of hybridoma cells in various serum-free media, implying the mitogenic effect of sericin is independent from media. We also demonstrated that sericin-S successfully induced the proliferation of CTLL-2, an established T lymphocyte cell line, under IL-2 starvation conditions. These results indicate that sericin, particularly sericin-S, improves serum-free mammalian cell culture.