使用不同胰酶消化PK15细胞对比试验

The PK15 cell comprison test digested with different pancreatic enzymes

为了寻找消化细胞温和且对细胞生长影响小的消化液,为生产疫苗提供更好细胞状态的PK15细胞,本试验使用0.25%胰酶A与0.25%胰酶B消化PK15细胞。两种胰酶均在常温中消化细胞且各分两组,一组消化完弃去消化液终止消化,另一组消化完不弃去消化液直接加入营养液终止,分种后进行培养观察。主要观察两种胰酶的消化终止时间、细胞分散效果、24 h细胞状态及两种胰酶消化时消化液弃去与不弃去的细胞状态。结果表明0.25%胰酶A消化PK15细胞,无论消化液弃去与不弃去,消化时间均为5~6 min,分散效果均较好且细胞状态都好。而0.25%胰酶B消化PK15细胞,消化时间均为1~2 min,消化液弃去,消化效果较好且细胞状态好;消化液不弃去,分散效果不好且细胞状态较差,生长较缓慢。消化PK15细胞时0.25%胰酶A较0.25%胰酶B更温和,0.25%胰酶A对细胞损伤较小。

In this work,0.25%pancreatin A and0.25%pancreatin B are used to digest PK15 cells so that a digestive juice which digest cells mildly and has little influence on the growth of cells is found.Simultaneously,this study can provide PK15 cells with better cellu鄄lar status for producing vaccines.Both of the types of pancreatic enzymes were digested under room temperatureand were divided into two groups.One group discard digestive juice to terminate digestion,another group didn't abandon digestive juice and add nutrient so鄄lution to terminate digestion,then culture and observe cells after dividing.The digestion time,cell dispersion,24 h cell status and sta鄄tus of cells with pancreatin which was keep and discard were main observed.The results show that when 0.25%pancreatin A was used to digest PK15 cell,whether discard or keep digestive juice,digestion time is 5~6 minutes and the dispersion effect is good.However,digestion time of PK15 cells by using 0.25%pancreatin B was 1~2 minutes.After discarding digestive fluid,digestion effect is good and the cells is in good condition.When digestive fluid was keep,the dispersion effect is not good,and growth of cells was poor and slow.This indicates that when digesting PK15 cells,0.25%pancreatin A is more mild than 0.25%pancreatin B and has less damage to cells.