BACKGROUND:
Lentivirus can infect divided and undivided
cells. It remains uncertain whether the
lentivirus can successful y infect primary ovarian
granulosa cells.
OBJECTIVE:
To investigate infecting ratio and cel
apoptosis of
lentivirus carrying bcl-2 gene in primary
human ovarian granulose
cells cultured
in vitro.
METHODS:
The lentiviral vector
carrying bcl-2 gene was constructed using
molecular biology, and packaged into
lentivirus with high titer. The resulting recombinant
lentivirus carrying bcl-2 genes were then used to infect primary
human ovarian
granulosa cells in vitro at different multiplicity of
infection, 10, 50, 100, 200, and 400.
Infection efficiency and cel proliferation were observed at 24, 48, 72, and 96 hours fol owing
infection. Cel
apoptosis was detected by
flow cytometry, and
bcl-2 gene transcription was assessed using
reverse transcription PCR. RESULTS AND
CONCLUSION:
Primary
human ovarian
granulosa cells adhered at 24 hours, and exhibited polygon- or fusiform-shape and colony-like
growth. When multiplicity of
infection was 100, cel appearance and
growth remained unchanged, and
infection efficiency was high, which reached the peak up to 72 hours. Moreover, the positive rate was up to 60% in
granulosa cells.
Lentivirus carrying bcl-2 gene could increase expression of Bcl-2
protein and inhibit
apoptosis of primary ovarian
granulosa cells.