Nayve F R, Motoki M, Matsumura M, Kataoka H
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki-ken, Japan.
Cytotechnology. 1991 Jun;6(2):121-30. doi: 10.1007/BF00373029.
Serum-free perfusion cultures of hybridoma TO-405 cells were carried out in spinner flasks coupled with zeolite A-3 packed beads. Ammonia was selectively removed from the culture broth by passing cell free permeate from ceramic cross flow filtration, through the zeolite packed bed. Ammonia concentration in the culture broth was effectively maintained between 1 to 4 mmol/l which was below the inhibitory concentration for cell growth. Maximum cell density levels of 10(7) cells/ml as well as improved percentage cell viability higher than in serum-supplemented cultures were feasible in this system. The possible effects of shear stress, generated by variation of the flow rates of the broth through the ceramic filter module, on the growth of the hybridoma cells were investigated. Backwashing, by reversing the direction of the permeate, was found necessary to prolong the life of the filter. Variation of the flow rates of the broth through the ceramic module between 0.29 m/s to 0.59 m/s did not cause immediate cell damage but growth was repressed at the higher flow rate. This study also showed that glutamine appears to be one of the factors limiting the growth of the hybridoma cells.
在装有A型3分子筛填充珠的转瓶中对杂交瘤TO - 405细胞进行无血清灌注培养。通过使陶瓷错流过滤产生的无细胞渗透液流经分子筛填充床，从培养液中选择性去除氨。培养液中的氨浓度有效维持在1至4 mmol/L之间，低于细胞生长的抑制浓度。在该系统中，可行的最高细胞密度水平为10(7)个细胞/ml，且细胞活力百分比高于补充血清的培养物。研究了培养液通过陶瓷过滤模块的流速变化产生的剪切应力对杂交瘤细胞生长的可能影响。发现通过使渗透液反向进行反冲洗对于延长过滤器寿命是必要的。培养液通过陶瓷模块的流速在0.29 m/s至0.59 m/s之间变化不会立即导致细胞损伤，但在较高流速下生长受到抑制。该研究还表明谷氨酰胺似乎是限制杂交瘤细胞生长的因素之一。
