ST贴壁细胞驯化悬浮及无血清培养工艺开发

Development of ST adherent cell domestication suspension and serum-free culture technology

[目的]将购自ATCC的ST贴壁细胞通过自主驯化,使其能在甘肃健顺生物科技有限公司(简称健顺生物)自产的无血清培养基CD ST 258中悬浮生长且能稳定传代,在反应器中能高密度生长。[方法]采用高血清贴壁培养转无血清悬浮培养;单因素实验优化氨基酸、维生素、无机盐等培养基组分;单因素实验优化生物反应器的pH、溶氧和转速三个参数。[结果]悬浮细胞在无血清培养基中传代,72 h细胞密度高于8.0×10^(6)cells/m L;反应器中细胞最高生长密度可达3.0×10^(7)cells/m L。[结论]ST贴壁细胞可驯化为悬浮细胞,能在无血清培养基中稳定传代且在一次性反应器中可高密度生长。

[Objective]To adapt ST adherent cells which are purchased from ATCC and make it possible to grow in the serum-free medium CD ST 258 developed by Biosciences Co.,Ltd.(short for JS Bio)independently.At the same time,it can be stable passage and high density growth in the bioreactor.[Method]The techniques including high-serum adherent culture to serum-free suspension culture;single-factor experiments to optimize medium components such as amino acids,vitamins,and inorganic salts;single-factor experiments to optimize the three parameters of bioreactor p H,dissolved oxygen and speed.[Result]It showed that the suspension cells were passaged in serum-free medium,and the cell density was higher than 8.0×10^(6) cells/mL at 72 hours;the highest cell growth density in the bioreactor could reach 3.0×10^(7) cells/mL.[Conclusion]ST adherent cells can be planted culture into suspension cells,can be passaged stably in serum-free medium and can grow at high density in single-use bioreactors.