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A Strategy to Identify Compounds that Affect Cell Growth and Survival in Cultured Mammalian Cells at Low-to-Moderate Throughput
Published on: September 22, 2019
Establishment of a low-tumorigenic MDCK cell line and study of differential molecular networks
Gui-Lan Ma1, Zi-Lin Qiao2, Dan He2
1College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, 730030, PR China; Gansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou, 730030, PR China.
Abstract:
Influenza is an acute respiratory infection caused by the influenza virus, and vaccination against influenza is considered the best way to prevent the onset and spread. MDCK (Madin-Darby canine kidney) cells are typically used to isolate the influenza virus, however, their high tumorigenicity is the main controversy in the production of influenza vaccines. Here, MDCK-C09 and MDCK-C35 monoclonal cell lines were established, which were proven to be low in tumorigenicity. RNA-seq of MDCK-C09, MDCK-C35, and MDCK-W73 cells was performed to investigate the putative tumorigenicity mechanisms. Tumor-related molecular interaction analysis of the differentially expressed genes indicates that hub genes, such as CUL3 and EGFR, may play essential roles in tumorigenicity differences between MDCK-C (MDCK-C09 and MDCK-C35) and MDCK-W (MDCK-W73) cells. Moreover, the analysis of cell proliferation regulation-associated molecular interaction shows that downregulated JUN and MYC, for instance, mediate increased proliferation of these cells. The present study provides a new low-tumorigenic MDCK cell line and describes the potential molecular mechanism for the low tumorigenicity and high proliferation rate.
Insights
New low-tumorigenic Madin-Darby canine kidney (MDCK) cell lines were developed for influenza vaccine production. These cell lines show reduced tumorigenicity and high proliferation rates, addressing key concerns in vaccine manufacturing.
Area of Science:
- Biotechnology
- Virology
- Cell Biology
Background:
- Influenza virus is a significant respiratory pathogen.
- Vaccination is the primary method for influenza prevention.
- Madin-Darby canine kidney (MDCK) cells are crucial for influenza virus isolation but have high tumorigenicity concerns for vaccine production.
Purpose of the Study:
- To establish and characterize novel, low-tumorigenic MDCK cell lines.
- To investigate the molecular mechanisms underlying tumorigenicity and proliferation in MDCK cells.
Main Methods:
- Development of monoclonal cell lines MDCK-C09 and MDCK-C35.
- RNA sequencing (RNA-seq) of MDCK-C09, MDCK-C35, and MDCK-W73 cells.
- Bioinformatic analysis of differentially expressed genes and molecular interactions.
Main Results:
- MDCK-C09 and MDCK-C35 cell lines exhibited significantly lower tumorigenicity.
- RNA-seq identified key genes (e.g., CUL3, EGFR) potentially involved in tumorigenicity differences.
- Downregulation of JUN and MYC was associated with increased cell proliferation.
Conclusions:
- Novel low-tumorigenic MDCK cell lines (MDCK-C09, MDCK-C35) have been successfully established.
- Potential molecular mechanisms for reduced tumorigenicity and enhanced proliferation have been elucidated.
- These findings offer a promising alternative for safer influenza vaccine production.
