Culture Vero cells

Fig. 2. Indirect immunofluorescence labeling of cell-bound component II. Component II (0.5ng/well) was incubated with cultured Vero cells at 37 °C for 7.5 min (a) not permeabilized with Triton X-IOO-PBS and (b) permeabilized with Triton X-100-PBS. For details see text (Section 9.5.4)... 

Lastly, these authors performed immunofluorescence microscopy (see section 4.3.4, Lectin histochemistry) to localize the PGase in cultured Vero cells. To allow for... 

Besides the role of retinoids in host immune defenses to viral infections (see [6] for review), retinoids may also affect virus replication. Angulo et al. [44] characterized three RA response elements in the promoter region of human cytomegalovirus (hCMV) and demonstrated the necessity for RAR and RXR in the viral promoter s positive response to RA. In contrast, the replication of herpes simplex virus-1 (HSV-1) in cultured Vero cells was inhibited by isomers of RA, but not retinol inhibition occurred without evident induction of IFN-a or IFN-p gene expression [45]. RA also protected HL-60 and WISH cells from infection with vesicular stomatitis virus (VSV) [46] however, in this case RA significantly increased the ability of IFN-a to decrease virus replication. These apparently contrasting effects of RA on hCMV as compared to HSV-1 or VSV replication further illustrate the potential for retinoids to act either positively or negatively in host resistance to viral infection and anti-viral immunity. Retinoid-IFN interactions are further discussed later in this chapter. 

Wang Y, Ouyang F (1999), Bead-to-bead transfer of Vero cells in microcarrier culture, Cytotechnology 31 221-224. 

Mendon a RZ, Ioshimoto LM, Mendon a RMZ, De Franco M, Valentini EJG, Be ak W, Pereira CA (1993), Preparation of human rabies vaccine in VERO cell culture using a microcarrier system, Braz. J. Med. Biol. Res. 26 1305-1317. 

Mendon a RZ, Prado JCM, Pereira CA (1999), Attachment, spreading and growth of VERO cells on microcarriers for the optimization of large scale cultures, Bioprocess Eng. 20 565-571. 

Mendon a RZ, Arrozio SJ, Antoniazzi MM, Ferreira JMC Jr, Pereira CA (2001), Metabolic active-high density VERO cell cultures on microcarriers following apoptosis prevention by galactose/glutamine feeding, J. Biotechnol. 97 13-22. 

Montagnon B, Vincent-Falquet JC, Fanget B (1984), Thousand litre scale microcarrier culture of VERO cells, Dev. Biol. Stand. 55 37-42. 

Nahapetian AT, Thomas JN, Thilly WG (1986), Optimization of environment for high density VERO cell culture effect of dissolved oxygen and nutrient supply on cell growth and changes in metabolites, J. Cell Sci. 81 65-103. 

Yokomizo AY, Antoniazzi MM, Galdino PL, Azambuja NJ, Jorge SAC, Pereira CA (2004), Rabies virus production in high Vero cell density cultures on macroporous microcarriers, Biotechnol. Bioeng. 85 506-515. 

Figure 7. Concentration profile of selected amino acids in Vero cell culture over time. The cell culture was sampled on days 4,6,8 (prior to feeding), 16 and 26. The culture was fed with initial media concentrate on days 4, 6 and 8, indicated by the arrows. At each time interval, an aliquot was deproteinized and derivatized according to the standard procedure. Amino acids were separated and quantified using HPLC System B and applying the optimized conditions for Mixture 11.

The Chinese cell culture-derived inactivated vaccine is produced in primary hamster kidney cells a highly purified Vero cell culture-derived inactivated vaccine is under clinical development in France. A Chinese live-attenuated vaccine is also produced in primary hamster kidney cells. The efficacy of one dose is 80%, and the efficacy of two doses, given 1 year apart, is 97.5% this vaccine was evaluated in a randomized trial in 26 239 children, half of whom received the vaccine and half served as controls, and its adverse effects have been reviewed (SEDA-22, 350). 

Originally cultures were established in Earle s BSS containing 0.5% lactalbumin hydrolysate, 0.1% yeast extract, 0.1% polyvinylpyrrolidone and 2-5% FBS but they can be maintained successfully in Eagle s MEM (EBSS) with added 7.5% bovine serum and 2.5% FBS or Medium 199 supplemented with 5% FBS. Vero cells are adaptable to batch and continuous perfusion culture. 

Van der Meer RX, Philippi MC, Romein B, Van der Velden de Groot CAM Beuvery EC (1993) Towards a strategy for high density cultures of Vero cells in stirred tank reactors. In Kaminogawa S, Ametani A Hachimura S (eds) Animal Cell Technology Basic and Applied Aspects, pp. 335-340. Kluwer Academic, Dordrecht, The Netherlands. 

In an alternative system, cells of the test culture can be inoculated onto coverslips preinoculated with an indicator cell line, such as the Vero African Green Monkey cell line. In this case the Vero cell should be inoculated at a cell concentration of 1 x 104 cells/mL and left for 4—24 h prior to addition of the test sample. The major advantage of this system, which overcomes the additional time required to set up, is the increased sensitivity achieved by the increased surface area of cytoplasm in Vero cells, which aids in revealing the mycoplasma. This system also enables the mycoplasma screening of serum and other reagents that can be inoculated directly onto the indicator cell line. 

Monolayers or African green monkey kidney (Vero) cells (for HSV) and human embryonic lung (HEL) fibroblasts (for VZV and CMV), grown in 25-cm2 tissue-culture flasks. 

Monolayers of Vero cells (for HSV) or HEL cells (for CMV and VZV) grown in 6-well tissue culture plates. 

Monolayers of Vero cells grown in 175-cm2 tissue culture flasks (Falcon). 

Use 8-10 tissue culture flasks (175-cm2) of Vero cells grown to confluence. 

However, the presence of cytotoxicity in the extract could be due to other toxins or bacterial products, it is necessary to perform a neutralization assay with specific antisera to confirm that the cytopathic effect is due to the production of Shiga toxins. Moreover, in absence of neutralization assay, VGA lacks specificity (Rahn et al. 1996). Therefore, the Vero Cells assay have been largely supplanted by immunologically-based and DNA-based method for the detection of STEC, but is still used for confirmation of Shiga toxin production from pure cultures. 

Solasonine inhibited larval development and pupation in Earias insulana [644], inhibited elongation of letuce seed radicles [646], inhibited the infectivity of herpes simplex virus type 1 and was cytotoxic to Vero cell cultures [652]. Solasonine weakly inhibited mycelium development in the fungus Phoma medicaginis synergism was observed in combination with 290 [647]. Solasonine lysed Penicillium notatum-derived protoplasts and bovine erythrocytes [648] and weakly disrupted stigmasterol and ergosterol liposomes [649] in each case, 290 was considerably more active. The completely assigned H and 13C NMR spectra for 291 have been reported [642],... 

Rabbit HCP-mix antiserum. The polyclonal rabbit anti-HCP serum was obtained by a cascade immunization of rabbits with a HCP mix from Vero cells. Three days before immunization blood was obtained from the rabbits. This pooled serum serves as pre-immune rabbit serum. The HCP mix, a blank culture of Vero cells is three times frozen and thawed to induce cell lysis. The culture supernatant was pooled and filtered. The pore size of the filters was comparable with those used during the IPV Vero production process. The HCP mix was divided in small aliquots, rapid frozen using dry ice and stored at -70 °C until use. 

With FMAU as a lead compound, the syntheses of other 5-alkyl substituted 2 -fluoro-ara-uracils were undertaken ( , ) including 1-(2 -deoxy-2 -f luoro-p-D-arab inofuranosyl )-5-e thy 1 uracil [FEAU]. As shown in Table I, though FEAU was approximately one log order less potent than FMAU against HSV-1 and HSV-2 infected Vero cells in culture, the former exhibited far less host cell toxicity, resulting in an extremely favorable therapeutic index (9.10). 

Treatment vas administered by i.v. injection twice daily beginning 48 hours after virus inoculation and continuing for ten days Viremia was determined by culture of lymphocytes collected from 3 mL of heparinized blood on Indicated days post-infection (p.i.). The mean plaque-forming units expressed is the average number of plaques present in two flasks of Vero cell co-cultures inoculated with each lymphocyte suspension. 

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