Giard D J, Fleischaker R J
Antimicrob Agents Chemother. 1980 Jul;18(1):130-6. doi: 10.1128/AAC.18.1.130.
Various parameters were examined for their effects on interferon production with human fibroblast cells (FS-4) grown on microcarriers, using a superinduction procedure. Optimal concentrations of cycloheximide and actinomycin D during induction were 10 micrograms/ml, respectively. Cells required 5 to 6 h of exposure to cycloheximide and at least 2 h of exposure to actinomycin D to achieve maximum yields. FS-4 cells were found to grow well at low serum concentrations (2.5 and 5%) and actually produced higher yields of interferon than cells grown at higher serum concentration (10 and 20%). Kinetics of interferon production at various temperatures revealed that significantly higher yields could be obtained at 34 degrees C than at either 37 or 30 degrees C. Priming cells for 16 h with 50 U of interferon per ml resulted in consistently higher yields of interferon. By modifying the superinduction procedure in accordance with the above findings, it is now possible to consistently obtain interferon yields of greater than 20,000 international units per 10(6) cells.
采用超诱导程序,研究了各种参数对微载体上生长的人成纤维细胞(FS-4)产生干扰素的影响。诱导过程中环己酰亚胺和放线菌素D的最佳浓度分别为10微克/毫升。细胞需要暴露于环己酰亚胺5至6小时,暴露于放线菌素D至少2小时才能达到最大产量。发现FS-4细胞在低血清浓度(2.5%和5%)下生长良好,实际上比在高血清浓度(10%和20%)下生长的细胞产生的干扰素产量更高。不同温度下干扰素产生的动力学表明,在34℃下可获得的产量明显高于37℃或30℃。用每毫升50单位的干扰素对细胞进行16小时的预处理,可使干扰素产量持续提高。根据上述发现修改超诱导程序后,现在能够始终如一地获得每10(6)个细胞超过20,000国际单位的干扰素产量。