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Production of a feline parvovirusvaccine using monolayer cell systems in roller flasks and microcarriers
Abstract:
A parvovirus-strain originating from a case of spontaneous panleukopenia has been adapted and propagated in a feline lung fibroblastic cell line. Cultivation of parvovirus infected monolayer cells was carried out in 11 glass flasks containing 50 ml media and on microcarriers in 0.5 - 101 stirring flasks with 0.5-61 media. Harvest of propagated virus from flask monolayer and microcarrier cultures was performed daily for 5 days during one week. The yields of virus antigen per/ml tissue culture media were equal in the two systems, as monitored by hemagglutination and ELISA assays. The seeding and harvesting procedures of virus were found to be simpler with the microcarrier system than with roller flasks. The scaling up is in progress and comparable results have been obtained in 101 scale.
Insights
This study adapted a feline panleukopenia virus strain in cell culture. Microcarrier systems offer simpler virus seeding and harvesting compared to traditional flask methods, with comparable yields.
Area of Science:
- Virology
- Cell Biology
- Veterinary Medicine
Background:
- Feline panleukopenia virus (FPV) is a significant pathogen in cats.
- Traditional cell culture methods for virus propagation can be labor-intensive.
- Developing efficient methods for FPV antigen production is crucial for diagnostics and vaccine development.
Purpose of the Study:
- To adapt and propagate a FPV strain in a feline lung fibroblastic cell line.
- To compare the efficiency of microcarrier-based cell culture with traditional monolayer flask cultures for FPV propagation.
- To evaluate virus antigen yield and procedural simplicity in both culture systems.
Main Methods:
- Adaptation and propagation of FPV in feline lung fibroblastic cells.
- Cultivation in both monolayer glass flasks and microcarrier-based stirred flasks.
- Daily virus harvest over 5 days for yield assessment.
- Quantification of virus antigen using hemagglutination and ELISA assays.
Main Results:
- Comparable virus antigen yields per ml of tissue culture media were achieved in both monolayer and microcarrier systems.
- Microcarrier cultures demonstrated simpler seeding and harvesting procedures compared to roller flasks.
- Successful scaling up to 10L scale with comparable results was achieved.
Conclusions:
- Microcarrier technology provides an efficient and simpler alternative for FPV propagation in cell culture.
- This method holds promise for scaled-up production of FPV antigen for research and diagnostic purposes.
- Further scaling up efforts are ongoing, indicating the potential for industrial application.