Study of Vero Cell Attachment and Growth in A Modified Serum-Free Medium (SFM01-M) Supplementing with Different Concentrations of Vitamin C

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Mancoh Posung, et al


New and reused tissue culture flasks (T-flasks) without precoating with adhesion factors (e.g. fibronectin, laminin, collagen) were used to subculture Vero cells for 3 passages (P1, P2, P3) by using a modified serum-free medium (SFM01-M). A comparison of the cell yields obtained from those passages of new or reused T-flask cultures was not significantly different. The effect of vitamin C on Vero cell growth showed that the cell yields obtained from SFM01-M supplemented with 25-50 mg/L of vitamin C significantly decreased in contrast to those obtained from a lower concentration. The analysis of secreted proteins using SDS-PAGE in SFM01-M supplemented with different concentrations of vitamin C (0, 10, 25, 50 mg/L) during the first 24 h of cultures showed that all cultures gave the similar protein bands with a molecular weight (MW) of approximately 146 kDa. This protein band specifically reacted with monoclonal anti-collagen type I using a direct enzyme-linked immunosorbent assay (ELISA). In microcarrier culture, the attachment of Vero cells on CytodexTM1 microcarriers could be observed as spherical shape during the first 3 h of culture. The maximum cell concentration was 14.90 ± 0.56 x 104 cells/mL at day 6 of culture. From this study, the attachment and growth of Vero cells on new and reused T-flasks including on CytodexTM1 microcarriers can be achieved by using SFM01-M.

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