rCHO细胞无血清适应及悬浮培养

Adaption and Suspension Culture of rCHO Cells Producing HBsAg in Serum-free Medium

考察了血清对rCHO(C28)细胞生长与产物(HBsAg)表达的影响,发现φ血清<0.01时细胞出现明显的代谢转换;考察了微量元素、氢化可的松、混合脂类与短多肽混合物(TL混合物)对细胞的影响,建立了适于rCHO(C28株)生长的无血清培养基MT-SFM;在将细胞从有血清转到无血清状态时,阶段性降血清比直接降血清更有利于rCHO(C28)细胞的无血清适应;当细胞适应了MT-SFM无血清培养基后,批培养中细胞的最大平均比生长速率可达0.65d-1,产物HBsAg的滴度在32～64之间。

Effects of different concentration of serum on the growth and HBsAg production of rCHO(C28) cells were studied, and the critical concentration of serum for the metabolic change of rCHO (C28) cells determined was φ__(NBS)<0.01. Effects of trace elements, hydrocortisone, lipid mixture and TL mixture on the growth of rCHO (C28) cells were also studied, and MT-SFM, a new serum-free media suited for rCHO (C28) cells, has been successfully developed. When rCHO (C28) cells were transferred from media with serum to serum-free media, the gradual adaptation was found to be more benificial than the direct adaptation. After rCHO (C28) cells have adapted to MT-SFM, the maximum of average specific growth rate was 0.65 d^(-1) and the titre of HBsAg was between 32 and 64 in batch culture.