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Stable Expression Stromal Cell-Derived Factor -1 Alpha Endothelial Cells Line Establishment and it’s Adhesion Activity with Monocyte

Chinese Journal of Arteriosclerosis(2007)

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摘要
Aim To study the effect of stromal cell derived factor-1 alpha (SDF-1α) on adherence of ECV304/THP-1. Methods Rat SDF-1α gene and pcDNA3.1 were both cut with EcoRⅠ, dephosphorylated, and connected in connecting buffer. Then, the connecting product was transformed to DH5α by calcium chloride, scanned by ampicillin, and cut by enzyme for the right inserting identification, and sub-selected by G418, SDF-1α expression of ECV304 was measured by reverse transcription polymerase chain reaction (RT-PCR) to get a stabilized SDF-1α expressional cells line. Trans-gene ECV304 was seeded in a six-pore plate. Then, THP-1 was added and incubated for 30 minutes, washed with PBS about 3 times, to remove unattached cells. Cells number was counted under microscope from 5 visual field -up, under, left, right and center. Results Stable SDF-1α expression cells line was obtained which was identified by RT-PCR (about 360 bp), and the adherence ability of trans-gene ECV304 was ten times and more than control, and CXCR4 mono-antibody obviously inhibited this adherence ability (P0.01). Conclusion Endogenous SDF-1α improves adhesion of ECV304/THP-1.
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关键词
endothelial cells,adhesion activity,cell-derived
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