Application Of Suspension Cell Culture For Preparation Of Viral Antigens

Introduction: HeLa cell is a widely studied cell line derived from human cerzix adenocarcinoma. The cells are epithelial - like in morphology and susceptible to polio viruses and adenovirus type 5. HeLa cells are usually as monolayer for isolation and propagation of some human viruses, as well as for the expression of recombinant proteins, including hepatitis B surface antigen. The aim of this study was to multiply the HeLa cells as suspension culture, and evaluate their susceptibility polio virus types 1, 2, 3 and adenovirus type 5.
Material and Methods: HeLa cells were grown in minimum essential medium supplemented with 10% heat-inactivated calf serum. HEPES buffer was used to obtain the best environmental condition for cell growth. To omit the calcium carboxy, methyl cellulose (CMC) and ethylene diamin tetraacetic (EDTA) were applied. After being adapted to suspension cell culture, cells were tested for their ability to host poliovirus types 1, 2 and 3 as well as adenovirus type 5,. Results of virus multiplication in both suspension and monolayer culture of the HeLa cells were compared.
Results: It was shown that there is close relation between the number of initiatins seed cells with doubling time of the cells. The best results were obtained when the number of initial seeds were 3x105 per ml for culturing in 96 well plates, and 4x105 for suspension culture . To see the effect of different culture conditions on cell growth and doubling time it was shown that CMC inhibited the cell growth while, reduced calcium concentration, HEPES buffer, and modified culture medium resulted in doubling of cells in about 48 hours. Both monolayer and suspension cell culture were equally susceptible to the viruses.
Conclusion: This study indicated that HeLa cells could be adapted to be culture as suspension, and different conditions either advantageous or disadvantageous to the cell growth and multiplication. The suspended cells maintained their susceptiblity to the test viruses. Using suspension culture of HeLa cells gives a great opportunity to use a much less volume and medium to obtain the same amount and titer of the desired virus.