Park Soonyong, Kim Ji Yul, Ryu Kyoung-Hwa, Kim Ah-Young, Kim Jaemun, Ko Young-Joon, Lee Eun Gyo
Bioprocess Engineering Center, KRIBB, 30 Yeongudanjiro Ochang-eup, Chungju-si 28116, Korea.
Department of Bioprocess Engineering, Korea University of Science and Technology (UST), Daejeon 34113, Korea.
Vaccines (Basel). 2021 May 13;9(5):505. doi: 10.3390/vaccines9050505.
The baby hamster kidney-21 (BHK-21) cell line is a continuous cell line used to propagate foot-and-mouth disease (FMD) virus for vaccine manufacturing. BHK-21 cells are anchorage-dependent, although suspension cultures would enable rapid growth in bioreactors, large-scale virus propagation, and cost-effective vaccine production with serum-free medium. Here, we report the successful adaptation of adherent BHK-21 cells to growth in suspension to a viable cell density of 7.65 × 10 cells/mL on day 3 in serum-free culture medium. The suspension-adapted BHK-21 cells showed lower adhesion to five types of extracellular matrix proteins than adherent BHK-21 cells, which contributed to the suspension culture. In addition, a chemically defined medium (selected by screening various prototype media) led to increased FMD virus production yields in the batch culture, even at a cell density of only 3.5 × 10 cells/mL. The suspension BHK-21 cell culture could be expanded to a 200 L bioreactor from a 20 mL flask, which resulted in a comparable FMD virus titer. This platform technology improved virus productivity, indicating its potential for enhancing FMD vaccine production.
幼仓鼠肾-21（BHK-21）细胞系是一种连续细胞系，用于繁殖口蹄疫（FMD）病毒以生产疫苗。BHK-21细胞依赖贴壁生长，不过悬浮培养能够实现其在生物反应器中的快速生长、大规模病毒繁殖以及使用无血清培养基进行具有成本效益的疫苗生产。在此，我们报告了贴壁的BHK-21细胞成功适应悬浮生长，在无血清培养基中第3天活细胞密度达到7.65×10个细胞/毫升。与贴壁的BHK-21细胞相比，适应悬浮培养的BHK-21细胞对五种细胞外基质蛋白的黏附性更低，这有利于悬浮培养。此外，一种化学成分明确的培养基（通过筛选各种原型培养基选定）即使在细胞密度仅为3.5×10个细胞/毫升时，也能使批次培养中的口蹄疫病毒产量增加。悬浮培养的BHK-21细胞可从20毫升培养瓶扩大至200升生物反应器，产生的口蹄疫病毒滴度相当。这种平台技术提高了病毒生产力，表明其在增强口蹄疫疫苗生产方面具有潜力。
