Abstract:

BACKGROUND: Gene transfer of exogenous cells should be done for convenient and accurate detection of exogenous cell survival in the body.
OBJECTIVE: To explore the gene transfer efficiency of bone marrow cells of BALB/C×C57BL F1 mice mediated by retroviral vector and stimulated with cytokines.
METHODS: The viral supernatants were harvested with PA317-GCGPXSN and detected with NIH3T3. Then gene transfer of experimental group was made on bone marrow cells from BALB/C×C57BL F1 mice as target cells which were stimulated by stem cell factor, interleukin 3, interleukin 6. The gene transfer efficiency was detected by flow cytometry and PCR. Negative group was not treated with gene transfer. Positive group used PA317-GCGPXSN cells as target cells.
RESULTS AND CONCLUSION: ①The viral titers were 1.9×10⁸ CFU/L. ②The gene transfer efficiency of the experiment group was 76.04%, whereas that of the control group was 0.63%, there was obviously statistical difference (P < 0.01). PCR amplification procured the specific NeoR gene fragments. Results verified that this method can transfer exogenous gene into bone marrow cell genome of BALB/C×C57BL F1 mice efficiently.