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Tumorigenicity decrease in Bcl-xL deficient MDCK cells ensuring the safety for influenza vaccine production
Jiahao Zheng1,2, Boran Li3, Lanxin Jia1,2
1National Engineering Technology Research Center for Combined Vaccines, Wuhan, China.
Abstract:
Madin-Darby canine kidney (MDCK) cells are the recognized cell strain for influenza vaccine production. However, the tumorigenic potential of MDCK cells raises concerns about their use in biological product manufacturing. To reduce MDCK cells' tumorigenicity and ensure the safety of influenza vaccine production, a B-cell lymphoma extra-large (Bcl-xL) gene, which plays a pivotal role in apoptosis regulation, was knocked-out in original MDCK cells by CRISPR-Cas9 gene editing technology, so that a homozygous MDCK-Bcl-xL-/- cell strain was acquired and named as BY-02. Compared with original MDCK cells, the proliferation and migration ability of BY-02 were significantly reduced, while apoptosis level was significantly increased, the endogenous mitochondrial apoptotic pathway were also modulated after Bcl-xL knock-out in MDCK cells. For tumor formation assays in nude mouse tests, all ten mice injected with original MDCK cells presented tumors growth in the injection site, in contrast to only one mouse injected with BY-02 cells presented tumors growth. These findings suggest that Bcl-xL knock-down is an effective strategy to inhibit tumor formation in MDCK cells, making BY-02 a promising genetically engineered cell strain for influenza vaccine production.
Insights
Researchers modified Madin-Darby canine kidney (MDCK) cells to create the BY-02 cell strain, significantly reducing tumorigenicity for safer influenza vaccine production. This engineered cell line shows reduced proliferation and increased apoptosis, enhancing vaccine safety.
Area of Science:
- Biotechnology
- Cell Biology
- Vaccinology
Background:
- Madin-Darby canine kidney (MDCK) cells are standard for influenza vaccine manufacturing.
- The inherent tumorigenic potential of MDCK cells poses a safety concern for biological product development.
Purpose of the Study:
- To engineer a safer MDCK cell line for influenza vaccine production by reducing tumorigenicity.
- To assess the impact of B-cell lymphoma extra-large (Bcl-xL) gene knockout on MDCK cell characteristics and tumor formation.
Main Methods:
- CRISPR-Cas9 gene editing was employed to knock out the Bcl-xL gene in MDCK cells, generating the BY-02 cell strain.
- Comparative analysis of proliferation, migration, apoptosis, and mitochondrial pathways between original MDCK and BY-02 cells.
- Tumor formation assays were conducted in nude mice using both cell types.
Main Results:
- The BY-02 cell strain exhibited significantly reduced proliferation and migration compared to original MDCK cells.
- Apoptosis levels were significantly increased in BY-02 cells, with modulation of the endogenous mitochondrial apoptotic pathway.
- Tumor formation assays demonstrated a substantial decrease in tumor incidence in mice injected with BY-02 cells versus MDCK cells.
Conclusions:
- Knocking down the Bcl-xL gene effectively inhibits tumor formation in MDCK cells.
- The genetically engineered BY-02 cell strain is a promising candidate for enhancing the safety of influenza vaccine production.
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