| Abstract |
A new cell line, designated BCIRL-PX2-HNU3, derived from 20
minced trypsinized pupae of diamond-back moth, Plutella xylostella, has
been established. Primary culture was set up in TC-199-MK medium on July
1, 1982. It was refed once a week. August 17, 48 days after explantation,
the first subculture was made just by flushing an equal volume of fresh
medium upon the confluent sheets, then the cells were dispensed into two
new culture flasks. The cells grew very rapidly. Since then they were
subcultured three times a week at a split ratio 1:2 and were subcultured
at a ratio 1:10 when they reached their passage 100. Now, this line is in
their 115 passage. BCIRL-PX2-HNU3 cells were not firmly attached and
floating cells were always observed in the medium. Most cells of this line
were round and oval in shape, some were elongated or spindle shaped,
although there were some other types of cells in the mixed cell population.
The population doubling time at 28 Celsius in 96 hours was 30 +- 2.06 and
the cell density reached, from 1x10^5 cells/ml inoculated, a level of
approximately 1.3x10^6 cells/ml after 8 days. Chromosomes were typical of
other lepidopteran cells. BCIRL-PX2-HNU3 was distinguished from cell lines
derived from other insect species by isoelectric focusing using the enzyme
phosphoglucoisomerase. Preliminary studies indicated that BCIRL-PX2-HNU3
cells could support the replication of some baculoviruses. The inclusion
bodies appeared in the nucleus of the cells within 48 hrs after
challenging the culture with Autographa californica's NPV. Inclusion
bodies of VHA-273, another heterologous baculovirus isolated from
Heliothis armigera in China could be replicated in Plutella cells as well,
either challenged the cells with infectious hemolymph or infectious
supernatant from infected cultures. One more interesting thing was that 6
days after challenging the cells with infectious hemolymph of its
homologous baculovirus, Plutella granulosis virus, GV capsules were found
in the cytoplasm of cultured Plutella cells, and it was confirmed by EM
pictures. But we failed to get it reconfirmed in subsequent experiments.
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