Poster presentation
187
GROWTH AND MAINTENANCE STUDIES OF
VERO CELL LINES
N. Tosheva, Sh. Khasanov, J. Abdurakhmanov, O. Ashirov, S. Sasmakov,
E. Yusupova, Sh. Azimova
S.Yu. Yunusov Institute of the Chemistry of Plant Substances Academy of sciences of the
Republic of Uzbekistan st. Mirzo-Ulugbek, 77, 100170 Tashkent
e-mail: genlab_icps@yahoo.com
Vero cells are derived from the kidney of the African green monkey and are one of
the most frequently used continuous mammalian cell
lines in microbiology and
molecular and cell biology research. Vero cells have been licensed in the United States
for production of both live (rotavirus, smallpox) and inactivated (poliovirus) viral
vaccines, and throughout the world Vero cells have been used
for the production of a
number of other viruses,
including Rabies virus, Reovirus and Japanese encephalitis
virus. The protocols outlined in this appendix detail procedures
for the routine growth
and maintenance of Vero cells in a research laboratory setting. In our study, we
investigated the growth conditions of Vero cell lines under laboratory conditions.
For
long-term storage, Vero cells are stored in liquid nitrogen or at -80°C. We
investigated the optimal conditions for growing Vero cells obtained from frozen stocks.
The vial (cryovial) of Vero cells was rapidly thawed by gently swirling in a 38°C
water bath. We transferred the Vero cell suspension from a cryovial to a 15 ml conical
tube containing 5 ml DMEM supplemented with FBS. Pelleted cells were centrifuged at
300 × g for 4 min at room temperature. The supernatant was removed and 5 ml of
DMEM supplemented with 10% FBS was added.
After freezing, Vero cells are best
recovered in a small (25 sm
2
or 50 sm
2
) tissue culture flask. We used 25 sm
2
flasks,
resuspended the cells in 5 ml medium. The flasks were incubated in an incubator at
37°C with 5% CO2. Cells are monitored daily. The media is changed every 2-4 days.
When cells reach more than 90% confluent monolayer, cells
are transferred to new
tissue culture flasks.
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