|
|
|
| WAVETM Bioreactors and Spinner Flasks: Comparison of the Process Performance of Microcarrier Bead to Bead Transfer and Scale up |
| LU Li-fang, SUI Li-li |
| Fast Trak China, GE Healthcare Lifesciences, Shanghai 201203, China |
|
|
|
Abstract Objective: Mammalian cell lines are widely used in biopharmaceuticals, such as the manufacturing of monoclonal antibodies, recombinant proteins and vaccines. For large scale adherent cell culture, microcarriers are developed and widely used in biopharmaceuticals. Adherent cell culture on microcarriers andbe done in both WAVETM reactors and stir tanks. However, to successfully scale up the microcarrier cultures, bead to bead transfer process is necessary. To develop bead to bead transfer, the new scale up technique, and evaluate the reliability of WAVETM reactor, a new generation of large scale fermenters, plenty of cell culture and bead to bead transfer experiments were done in WAVETM reactors and spinner flasks. The data were collected and analyzed. Method: Vero cells were inoculated and cultured with microcarrier Cytodex 1 in WAVETM reactors and spinner flasks, respectively. For good cell propagation, the culture conditions such as pH, temperature and nutrients were controlled appropriately. The cells grown confluent on the microcarriers were separated through bead to bead transfer process including wash, trypsin digesting, and re-inoculating, to be transferred a new scaled up culture. The efficiencies of bead to bead transfer process were evaluated through recording and analyzing the cell recoveries during wash and digestion, and cell growth viabilities after re-inoculum. Results: Statistical analysis showed that the average of cell recoveries of the bead to bead transfer processes from WAVETM reactors and spinner flasks were 67.56% and 39.39%, respectively. P value was below 0.000 3. The average inoculums re-attach efficiency from WAVETM reactors and spinner flasks were 95.17% and 78.45%, respectively. P value was 0.010 7. Conclusion: Bead to bead transfer process done in WAVETM reactors got much better performance in terms of cell recovery and cell viability, comparing with that in spinner flasks. Vero cells grown in WAVETM are in good conditions. As either seed train or production tank, WAVETM reactors wound be muchsuperior comparing with stir tanks.
|
|
Received: 04 June 2012
Published: 25 September 2012
|
|
|
|
|
[1] Kistner O, Howard MK, Spruth M, et al.Cell culture (Vero) derived whole virus (H5N1) vaccine based on wild-type virus strain induces cross-protective immune responses. Vaccine,2007, 25(32): 6028-6036.
[2] Genzel Y, Behrendt I, König S, et al.Metabolism of MDCK cells during cell growth and influenza virus production in large-scale microcarrier culture. Vaccine,2004, 22(17-18): 2202-2208.
[3] Trabelsi K, Majoul S, Rourou S, et al.Development of a measles vaccine production process in MRC-5 cells grown on Cytodex1 microcarriers and in a stirred bioreactor. Appl Microbiol Biotechnol,2012,93(3): 1031-1040.
[4] Microcarrier Cell culture-Principles and Methods. GE Healthcare Handbook, #18114062AB.
[5] Velez J O, Russell B J, Hughes H R, et al. Microcarrierculture of COS-1 cells producing Japanese encephalitis and dengue virus serotype 4 recombinant virus-like particles. J Virol Methods, 2008, 151(2):230-236.
[6] Liu C C, Lian W C, Butler M, et al. High immunogenic enterovirus 71 strain and its production using serum-free microcarrier Vero cell culture. Vaccine,2007, 25(1):19-24.
[7] Lupberger J, Mund A, Kock J, et al. Cultivation of HepG2.2.15 on Cytodex-3: higher yield of hepatitis B virus and less subviral particles compared to conventional culture methods.J Hepatol, 2006, 45(4):547-552.
[8] Bleckwenn N A, Bentley W E, Shiloach J, et al. Evaluation of production parameters with the vaccinia virus expression system using microcarrier attached HeLa cells.Biotechnol Prog, 2005, 21(2):554-561.
[9] Sun L Y, Lin S Z, Li Y S, et al. Functional Cells Cultured on Microcarriers for Use in Regenerative Medicine Research. Cell Transplantation, 2011, 20(1):49-62.
[10] Kistner O, Barrett P N, Mundt W, et al. A novel mammalian cell (Vero) derived influenza virus vaccine: Development,characterization and industrial scale production. Wiener Klinische Wochenschrift,1999, 111(5):207-214.
[11] Hundt B, Best C, Schlawin N, et al. Establishment of a mink enteritis vaccine production process in stirred-tank reactor and Wave Bioreactor microcarrierculture in 1-10 L scale.Vaccine, 2007, 25(20):3987-3995.
[12] 张立,严春,范卫民,等.Vero细胞的微载体培养——放大过程中的接种工艺. 华东理工大学学报,1998, 24(6):659-663. Zhang L, Yan C, Fan W M, et al. Inoculum Technology in Large-scaleCell Culture onMicrocarriers. Journal of East China University of Science and Technology, 1998, 24(6):659-663. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
