牛环形泰勒虫血源细胞的生物反应器培养——营养物质消耗及代谢产物积累对细胞生长繁殖的影响

The Culture of Bovine White Blood Cells Infected with Schizonts of Theileria Annulata in Bioreacter——Effects of Nutrients and Metabolites on the Growth of the cell culture

本文探讨了牛环形泰勒虫血源细胞活疫苗生产工艺中,营养物质消耗及代谢产物积累对血源细胞在生物反应器批培养中生长的影响。结果表明,牛环形泰勒虫血源细胞在生物反应器中接种5.0×105cells/ml,经60h细胞培养密度达到最大值2.19×106cells/ml。接种后在48h内,细胞生长迅速,未经生长适应期,就直接进入对数生长期;但培养到36h,葡萄糖就基本耗尽,48h后氨浓度上升,使得细胞生长的稳定期非常短暂,仅有12h;随后,细胞培养密度开始下降;96h,氨的积累量达到峰值270.52mg/l,细胞的生长受到明显的抑制,活性迅速下降,细胞生长进入衰亡期。到168h,细胞活力仅有5.91%。可见本次批培养中,营养物质葡萄糖初始浓度(2017.67mg/L)不足,而谷氨酰胺初始浓度可能(2.6mmol/L)偏高。

The effects of nutrients and metabolites on the growth of bovine white blood cells infected with schizonts of Theileria annulata in bioreactor of the production of live chizont vaccine,were studied. In bioreactor ,the condition of cell culture is that ,PH is 7.2-7.4,Temprature is 37℃, 5%CO2,95%Air ,and glucose is 2017.67 mg/L, L-glutamine is 6.0 mmol/L. It was found that, the cells growth from inoculum size （5.0× 10^5cells/ml） to maximal viable cell density （21.9×10^6eells/ml） in the period of 60 hours. The growth curve is as follows： no lag phase, logarithmic phase from 0 to 48 hours, stationary phase from 48 to 60 hours, and decline phase from 96 to 228 hours. But in 36 hours, the consumption of glucose is declined with sharpnees linearization and on 36 hours its concentration is nearly zero, while the concentration of ammonia increased, made the stationary phase was very short, only 12 hours. Then viable cell density was declined with slowness. On 96 hours, the concentration of ammonia reached to maximal was 270.52mg/L and decline phase started .On 168 hours, the viable cell was 5.19%.It suggested that, the concentration of Glucose was lower but Lglutamine was higher.