Domestication and Biological Characteristics of a Vero Cell line in Suspension Culture

Lanbo FU; Chaolie RUAN; Changxu CHEN; Yu GAO; Xiaolin BAO; Bochuan LIU; Jian ZHOU; Weidong LI

Article Contents

Article Navigation > Journal of Kunming Medical University > 2024 >

Lanbo FU, Chaolie RUAN, Changxu CHEN, Yu GAO, Xiaolin BAO, Bochuan LIU, Jian ZHOU, Weidong LI. Domestication and Biological Characteristics of a Vero Cell line in Suspension Culture[J]. Journal of Kunming Medical University.

Citation:

PDF( 2685 KB)

Domestication and Biological Characteristics of a Vero Cell line in Suspension Culture

1.
Institute of Medical Biology，Chinese Academy of Medical Science，Kunming Yunnan，650118
2.
State Key Laboratory Conservation and Utilization of Bio-Resources in Yunnan，Yunnan University，Kunming Yunnan，650091，China

Received Date: 2024-02-09

Abstract

Abstract

Objective To domesticate adherently cultured Vero cells into suspension-cultured cells and investigate their biological characteristics. Methods Cell culture was optimized using serum-free media and the "shaker adaptation method." Additionally, tumorigenicity, short tandem repeat （STR） profiles, morphological characters under transmission electron microscope （TEM）, and influenza virus sensitivity were analyzed in these cells. Results Suspension cell line Vero-S was successfully established. The growth period of Vero-S cells ranged from 24 to 72 hours, with a plateau phase between 108 and 144 hours, and a decline phase after 144 hours. The population doubling time （PDT） was 36 hours, and cell density ranged from 14.01 to 14.88 × 10⁸ cells/L. Average cell roundness ranged from 0.73 to 0.75, and cell diameter ranged from 12.12 to 14.44 µm. Vero-S cells formed tumors subcutaneously in nude mice, while Vero cells before suspension culture did not exhibit tumorigenicity. STR analysis indicated a primate origin of the cell line. Under electron microscopy, cell structure appeared intact. Vero-S cells showed insensitivity to influenza virus infection. Conclusion The established suspension cell line Vero-S exhibiting growth patterns similar to adherent cells, with uniform cell size and high-density growth supported by the established suspension culture medium. STR results demonstrated complete genetic stability of suspended Vero cells, with no apparent differences observed under transmission electron microscopy compared to adherent Vero cells. This study provides reference for suspension culture domestication techniques and can be applied in mechanistic studies of cell tumorigenicity.
- Vero cells,
- Cell suspension domestication technology,
- Tumorigenicity,
- Virus sensitivity

FullText(HTML)

References(25)

References

[1]	Ahmad N,Nawi A M,Jamhari M N,et al. Post-exposure prophylactic vaccination against rabies: A systematic review[J]. Iran J Public Health,2022,51(5):967-977.
[2]	Hu L,Sun J,Wang Y,et al. A review of inactivated COVID-19 vaccine development in China: Focusing on safety and efficacy in special populations[J]. Vaccines,2023,11(6):1045. doi: 10.3390/vaccines11061045
[3]	Wang L,Li J,Mu Q,et al. Study on immune persistence of the CTN-1V strain rabies vaccine in humans[J]. Journal of Virus Eradication,2024,10(1):100365. doi: 10.1016/j.jve.2024.100365
[4]	Liang C,Hui N,Liu Y,et al. Insights into forsythia honeysuckle (Lianhuaqingwen) capsules: A Chinese herbal medicine repurposed for COVID-19 pandemic[J]. Phytomed Plus,2021,1(2):100027. doi: 10.1016/j.phyplu.2021.100027
[5]	Someya K,Okemoto-Nakamura Y,Kurata T,et al. Establishment of measles virus receptor-expressing Vero cells lacking functional poliovirus receptors[J]. Microbiol Immunol,2023,67(3):166-170. doi: 10.1111/1348-0421.13047
[6]	Todesco H M,Gafuik C,John C M,et al. High-titer manufacturing of SARS-CoV-2 Spike-pseudotyped VSV in stirred-tank bioreactors[J]. Mol Ther Methods Clin Dev,2024,32(1):101189. doi: 10.1016/j.omtm.2024.101189
[7]	Guo M,Zheng R,Wu H L,et al. Inhibition of enterovirus 71 infection by polysaccharides extracted from Picochlorum sp. 122 via the AKT and ATM/ATR signaling pathways[J]. Arch Virol,2021,166(12):3269-3274. doi: 10.1007/s00705-021-05229-1
[8]	赵彩红,王美皓,李自良,等. 无血清悬浮培养MDCK细胞系的建立及生物反应器高密度培养[J]. 中国生物制品学杂志,2021,34(11):1362-1369.
[9]	刘丽,郭骐源,郝建丽,等. 无血清悬浮MDCK细胞培养流感病毒纯化工艺的初步建立[J]. 中国生物制品学杂志,2021,34(9):1100-1104.
[10]	李政蓉,金宏丽,黄培,等. BHK-21细胞悬浮培养狂犬病病毒(rCVS-11-dG株)工艺的建立及病毒免疫原性评价[J]. 中国生物制品学杂志,2021,34(2):129-134.
[11]	陈丽,倪敏舒,徐悦,等. 伪狂犬病毒在BHK-21悬浮细胞中的增殖特性[J]. 江苏农业科学,2022,50(10):166-170.
[12]	Göbel S,Pelz L,Silva C A T,et al. Production of recombinant vesicular stomatitis virus-based vectors by tangential flow depth filtration[J]. Appl Microbiol Biotechnol,2024,108(1):240. doi: 10.1007/s00253-024-13078-6
[13]	Cui W,Liu S. Optimization of adaptation parameters from adhesion cell culture in serum-containing media to suspension in chemically defined media by superlative box design[J]. Cytotechnology,2024,76(1):39-52. doi: 10.1007/s10616-023-00596-w
[14]	Du Q,Zhang X,Wang T,et al. Effects and mechanisms of animal-free hydrolysates on recombination protein yields in CHO cells[J]. Appl Microbiol Biotechnol,2022,106(22):7387-7396. doi: 10.1007/s00253-022-12229-x
[15]	Huang Z,Habib A,Zhao G,et al. CRISPR-Cas9 mediated stable expression of exogenous proteins in the CHO cell line through site-specific integration[J]. Int J Mol Sci,2023,24(23):16767. doi: 10.3390/ijms242316767
[16]	国家药典委员会编. 中华人民共和国药典三部 2020年版 [M]. 北京: 中国医药科技出版社,2020: 11-13.
[17]	Kiesslich S,Kim G N,Shen C F,et al. Bioreactor production of rVSV-based vectors in Vero cell suspension cultures[J]. Biotechnol Bioeng,2021,118(7):2649-2659. doi: 10.1002/bit.27785
[18]	Kim J,Kwon E J,Kim Y J,et al. Epigenetic repression of CHCHD2 enhances survival from single cell dissociation through attenuated Rho A kinase activity[J]. Cell Mol Life Sci,2024,81(1):38. doi: 10.1007/s00018-023-05060-8
[19]	Wu K,El Zowalaty A E,Sayin V I,et al. The pleiotropic functions of reactive oxygen species in cancer[J]. Nat Cancer,2024,5(3):384-399. doi: 10.1038/s43018-024-00738-9
[20]	Sato Y,Bando H,Di Piazza M,et al. Tumorigenicity assessment of cell therapy products: The need for global consensus and points to consider[J]. Cytotherapy,2019,21(11):1095-1111. doi: 10.1016/j.jcyt.2019.10.001
[21]	Yang Z,Yu S,Xu Y,et al. The screening and mechanism of influenza-virus sensitive MDCK cell lines for influenza vaccine production[J]. Diseases,2024,12(1):20. doi: 10.3390/diseases12010020
[22]	Ulengin-Talkish I,Parson M A H,Jenkins M L,et al. Palmitoylation targets the calcineurin phosphatase to the phosphatidylinositol 4-kinase complex at the plasma membrane[J]. Nat Commun,2021,12(1):6064. doi: 10.1038/s41467-021-26326-4
[23]	Logan M,Rinas K,McConkey B,et al. Vero cells gain renal tubule markers in low-calcium and magnesium chemically defined media[J]. Sci Rep,2022,12(1):6180. doi: 10.1038/s41598-022-10221-z
[24]	V á zquez-Ram í rez D,Jordan I,Sandig V,et al. High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system[J]. Appl Microbiol Biotechnol,2019,103(7):3025-3035. doi: 10.1007/s00253-019-09694-2
[25]	Kalle. S R M Z H. Adaptation of Vero cells to suspension growth for rabies virus production in different serum free media[J]. Vaccine,2019,37(47):6987-6995. doi: 10.1016/j.vaccine.2019.05.092

Relative Articles(20)

Relative Articles

[1]	Jun HE, Xuezhong GU, Yating LIN, Fan LI. Research Progress on the Mechanism of Regulatory T Cells in Tumor Microenvironment of Diffuse Large B-cell Lymphoma. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20230607
[2]	Shibing QIAN, Lingpeng ZHANG, Lingyun YIN, Changquan LI, Hu LI, Hongbin YU. Study on Osteogenic Ability of Dental Pulp Stem Cells at Different Concentrations. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20230214
[3]	Qingzhu ZHOU, Zhuo GONG, Xian ZHAO, Boyan LIU, Wenli HUANG, Jia HE, Liu LIU, Pufei PU. Recombinant Human Epidermal Growth Factor on the Repairing Effect of Sensitive Skin in High Altitude Areas in Yunnan Province. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20220627
[4]	Xifang WU, Yong WANG, Lisha PENG, Jie LUO, Feng WANG. Effect of VEGF-C on Radiation Sensitivity of Nasopharyngeal Carcinoma CNE-2 Cell Transplanted Mice. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20221205
[5]	Fan CAO, Yang-wen LIU, Lei WEI, Nan XIE, Yi-jian YANG, Cheng QIAN, Chun-mang LI, Bi-mang FU, Jiang-pin XU, Rong-ji YAN, Wang ZENG. Correlation between Hepatitis B Virus and Microvascular Invasion in Hepatocellular Carcinoma. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20210331
[6]	Hong-min DUAN, Nan CHEN, Yong-yan YANG, Yun-xia LI, Jin-xiang FENG, Hang LI, Xiu-wen HUANG, Dong-jin LV, Ming ZHANG, Yu-tao ZHAO. Effects and Mechanism of Anlotinib on Radiosensitivity of Non-small Cell Lung Cancer Cell Lines A549. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20210808
[7]	Yang Can , Gong Yu Hang , Wang Hai Feng , Li Hai Hao , Liu Jing Yu , Wang Wei , Wang Jian Song , Zuo Yi Gang , Chen Jian , Zhan Hui , Ding Ming Xia . The Research of Monoclonal Antibody KMP1 Inhibited Bladder Cancer EJ Cell Lines Growth and Metastasis in Vivo. Journal of Kunming Medical University,
[8]	Cui Kun Hua , Li Qi . Vitamin D Receptor Gene TaqI Polymorphisms and Steroid Responsiveness in Childhood Nephrotic Syndrome. Journal of Kunming Medical University,
[9]	Li Ying Fu . . Journal of Kunming Medical University,
[10]	Ai Yi Qin . . Journal of Kunming Medical University,
[11]	Wei Na . . Journal of Kunming Medical University,
[12]	Li Ying Fu . . Journal of Kunming Medical University,
[13]	Ai Yi Qin . . Journal of Kunming Medical University,
[14]	Qi Zong Ze . . Journal of Kunming Medical University,
[15]	Chen Shao Chun . . Journal of Kunming Medical University,
[16]	Zhao Chun Fang . . Journal of Kunming Medical University,
[17]	Chen Rui Bin . . Journal of Kunming Medical University,
[18]	Zhang Yan . . Journal of Kunming Medical University,
[19]	Yang De Biao . . Journal of Kunming Medical University,
[20]	. Clinincal and Pathological Analysis of Maxillomandibular Inflammatory Myofibroblastic Tumor. Journal of Kunming Medical University,