Abstract
Follicle-stimulating hormone (FSH) was produced in Chinese hamster ovary (CHO) cells using a perfusion bioreactor. Perfusion culture at 37 degrees C yielded a high cell density but a low FSH production. To investigate the effect of culture temperature in the range of 26-37 degrees C on cell growth and FSH production, batch cultures were performed. Lowering culture temperature below 32 degrees C resulted in growth suppression. However, specific productivity of FSH, q (FSH), increased as culture temperature decreased, and the maximum q (FSH) of 43.4 ng/10(6) cells/h was obtained at 28 degrees C, which is 13-fold higher than that at 37 degrees C. Based on the results obtained from batch cultures, we performed perfusion cultures with two consecutive temperatures. CHO cells were grown up to 3.2 x 10(7) cells/ml at 37 degrees C and culture temperature shifted down to 28 degrees C to obtain a high FSH titer. Soon after the maximum FSH titer of 21 mug/ml was achieved, a rapid loss of not only viable cell concentration but also cell viability was observed, probably due to the low activities of enzymes related to cell growth. Thus, the extension of production period at 28 degrees C is critical for the enhancement of FSH production, and the use of antiapoptotic genes seems to be promising.
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