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制备作为促有丝分裂因子的丝蛋白丝胶以优化哺乳动物细胞培养

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

作者信息

Terada Satoshi, Sasaki Masahiro, Yanagihara Kana, Yamada Hideyuki

机构信息

Department of Applied Chemistry and Biotechnology, University of Fukui, Japan.

出版信息

J Biosci Bioeng. 2005 Dec;100(6):667-71. doi: 10.1263/jbb.100.667.

Abstract

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.

摘要

我们之前报道过,分子量在5至100 kDa之间的小丝胶蛋白(sericin-S)是一种用于加速细胞增殖的细胞培养补充剂。在本研究中,我们考察了一种丝胶蛋白的新制备方法以及丝胶蛋白的几种应用。在非水解条件下制备的、分子量在50至200 kDa之间的大丝胶蛋白也能加速细胞增殖,但其效果不如sericin-S。我们还测试了其他不同程度水解的、具有各种分子量的丝胶蛋白制剂,但它们均未显著优于sericin-S,几种重组丝胶蛋白肽也同样如此。sericin-S在各种无血清培养基中均成功加速了杂交瘤细胞的增殖,这意味着丝胶蛋白的促有丝分裂作用与培养基无关。我们还证明,在白细胞介素-2饥饿条件下,sericin-S成功诱导了已建立的T淋巴细胞系CTLL-2的增殖。这些结果表明,丝胶蛋白,尤其是sericin-S,可改善无血清哺乳动物细胞培养。

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