Department of Applied Chemistry and Biotechnology, Fukui University, Bunkyo, Fukui, Japan, 776 27 8747),
Cytotechnology. 2002 Nov;40(1-3):3-12. doi: 10.1023/A:1023993400608.
Sericin, a constituent of the silkworm cocoon, was added to the culture of four mammalian cell lines: murine hybridoma 2E3-O,human hepatoblastoma HepG2, human epithelial HeLa and human embryonal kidney 293 cells. The proliferation of all cell lineswas accelerated in the presence of sericin. The hybridoma cellline was further studied. The 2E3-O cell line was so well adapted to serum-free medium that both the proliferation rate and maximum cell density in serum-free ASF103 medium were higher than in RPMI medium supplemented with all lots of FBS tested, and this proliferation was stimulated by the addition of sericin in a dose-dependent manner. Stimulation was observed at sericin concentrations from 0.01 to 0.1 %, although 1% sericin was severely harmful to the culture. In comparison with bovine serum albumin (BSA), a widely used supplement in serum-free medium, sericin had an equivalent effect on the proliferation of the hybridomas and sericin additively stimulated the proliferation with BSA. Although heat easily denatures and inactivates most proteins, the activity of sericin was not affected by autoclaving. In a similar manner to the silkworm-derived sericin, recombinant sericin synthesized in E. coli also stimulated the hybridoma proliferation, irrespective of whether it was autoclaved or filtered. Since BSA is obtained from bovine serum and the risk of infections such as bovine spongiform encephalopathy cannot be eradicated, sericin derived from insects could be a preferable culture medium supplement for stimulating the proliferation of mammalian cells.
丝胶是蚕茧的组成部分,被添加到四种哺乳动物细胞系的培养中:鼠杂交瘤 2E3-O、人肝癌 HepG2、人上皮 HeLa 和人胚肾 293 细胞。所有细胞系的增殖在丝胶存在的情况下都加速了。进一步研究了杂交瘤细胞系。2E3-O 细胞系非常适应无血清培养基,以至于在无血清 ASF103 培养基中的增殖率和最大细胞密度都高于补充了所有测试批次 FBS 的 RPMI 培养基,并且这种增殖以剂量依赖的方式被丝胶刺激。在丝胶浓度为 0.01%至 0.1%时观察到刺激,尽管 1%的丝胶对培养物有严重的危害。与广泛用于无血清培养基中的牛血清白蛋白 (BSA) 相比,丝胶对杂交瘤的增殖具有相同的作用,并且丝胶与 BSA 一起添加可刺激增殖。尽管热容易使大多数蛋白质变性和失活,但丝胶的活性不受高压灭菌的影响。以与蚕衍生的丝胶类似的方式,在大肠杆菌中合成的重组丝胶也刺激了杂交瘤的增殖,无论其是否经过高压灭菌或过滤。由于 BSA 是从牛血清中获得的,并且不能消除牛海绵状脑病等感染的风险,因此来自昆虫的丝胶可能是刺激哺乳动物细胞增殖的更可取的培养基补充剂。