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一株MDBK细胞的低血清悬浮驯化研究
王磊,杨承槐,刘莹
0
(中国兽医药品监察所)
摘要:
本研究选取一株背景清晰、纯净的MDBK贴壁细胞,通过直接驯化和逐级降低血清相结合的办法,获得了一株可以在低血清培养基中全悬浮培养的MDBK细胞。该细胞在含0.5%胎牛血清的培养基中悬浮培养48 h后的细胞密度稳定在5.0×106 cells/mL以上,培养72h后细胞密度最高可达1.16×107cells/mL,且细胞形态良好,活力在93%以上,具有良好的传代稳定性。对筛选获得的悬浮培养MDBK细胞株分别接种伪狂犬病毒和牛传染性鼻气管炎病毒后,检测病毒敏感性。结果发现细胞在24小时均发生了皱缩,72小时大部分死亡,呈碎片状。传染性牛鼻气管炎病毒滴度在24 h达到最大值,TCID50为10-7.6/ml;伪狂犬病毒滴度在48 h达到最大,TCID50为10-7.6/ml,说明虽然培养方式发生了改变,并没有影响该细胞对两种病毒增殖特性。本研究希望通过对MDBK细胞的全悬浮驯化研究,为相关病毒类疫苗规模化生产及工艺的升级改进提供细胞学基础资料。
关键词:  全悬浮培养、MDBK细胞、低血清、细胞驯化
DOI:
投稿时间:2020-09-21修订日期:2021-03-23
基金项目:中国兽医药品监察所所级课题(201808);
Domestication of a MDBK Cell Line to Suspension Culture by Low- Serum Medium
(China Institute of Veterinary Drug Control)
Abstract:
In this study, a well-defined and pure adherent bovine kidney cells(MDBK) was acclimated to the suspension condition though direct reducing serum and gradually reducing serum. The results showed that the MDBK cell density proliferated to 5×106 cells/mL after 48 h of suspension culture in low serum medium, and reached highest cell density to 1.16×107 cells/mL after 72 h with good morphology, viability bove 93%, well generational stability. In order to test virus sensitivity, the suspension MDBK cell lines were inoculated with pseudorabies virus (PRV) and bovine infectious rhinotracheitis virus (IBRV). The results showed that the cells were all shrunk in 24 h and most of them died in flakes at 72 h. The titer of IBR reached the maximum value at 24 h, TCID50 was 10-7.6/ml; the titer of PRV reached the maximum at 48 h, TCID50 was 10-7.6/ml. The result indicated that although the culture mode had changed, it did not affect the proliferation characteristics of the two viruses. We hope that the full suspension domestication of MDBK cells study can provides basic cytological data for scale production of related viral vaccines.
Key words:  suspension  MDBK cells  low serum  cell domestication  

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