Study of Vero Cell Growth in A Modified Serum-Free Medium (SFM01-M)

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Published: Jan 30, 2017
Keywords:
a modified serum-free medium microcarrier Vero cells

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Manoch Posung, et al
Medical Biotechnology Center Medical Life Science Institute Department of Medical Sciences Ministry of Public Health

Abstract

The aim of this work was to study a modified serum-free medium (SFM01-M) in supporting Vero cell growth in both static and microcarrier cultures. SFM01-M was developed using a systematic design of experiments (DoE). In static cultures, a concentration of rhEGF in SFM01-M was first studied in order to find its concentration suitable for using in the long-term subcultures of Vero cells and in an attempt to assess the minimizing of a cost of SFM01-M as a result of the addition of rhEGF. The results shown here indicated that the addition of rhEGF at least 0.015 mg/L in SFM01-M had effect on cell yields, particularly during subcultures under a serum-free condition (P1-P3). Additionally, a comparison of cell yields obtained from P3 between the rhEGF-added SFM01-M and the rhEGF-free SFM01-M cultures was found to be significantly different (P-value < 0.05). Therefore, in the long-term subcultures of Vero cells, a concentration of rhEGF at 0.015 mg/L was chosen to add to SFM01-M. The results showed that the cell yields obtained during P1-P9 were in the range of 7.09±0.12–7.33±0.33x105 cells/well (6-well plate) and then decreased gradually from P10 to P12. With the gradual decrease of growth ability, the cell yields obtained during P12-P20 were rather lower (5.75±0.39–5.96±0.36x105 cells/well) than those obtained from previous passages. By increasing an initial cell density of 4x104 cells/cm2, therefore, the cell yields could be increased in the range of 7.54±0.19–8.00±0.15x105 cells/well during P10-P20. In microcarrier cultures, the ability of Vero cells to attach and grow on CytodexTM1 solid microcarriers in SFM01-M in a spinner flask was found to be close to that of VP-SFM. From this study, Vero cells can be grown in SFM01-M in both static and microcarrier cultures. Moreover, a cost of SFM01-M is relatively cheap (approximately 70 US dollar per liter) and SFM01-M can also be an economical alternative for Vero cell cultures under a serum-free condition.

Article Details

Issue
Vol. 8 No. 8 (2016): JANUARY 2016 - DECEMBER 2016
Section
บทความวิจัย

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