OBJECTIVETo establish the co-culture model of human macrophage cell line (U937) with human vein umbilical cell line (ECV304), and to explore the feasibility of using concanavalin A (ConA) as U937 cell stimulator in regulating angiogenesis.

METHODSECV304 cells were cultured in vitro, and to which were respectively added U937 cells (1 x 10(5)), 25 microg/mL ConA, and U937 cell (1 x 10(5)) + ConA (25 microg/mL) after cell fusion rate reaching 60%, and then co-cultured for 48 hours. ECV 304 cells in conventional culture were used as controls. 3H-TdR incorporation test was employed to determine the DNA synthesis of vascular endothelial cells. Flow cytometry was used to determine the changes in the cell cycle, and RT-PCR was adopted to determine the expression of homeobox (HOXB2) mRNA.

RESULTSAfter conA stimulation to ECV 304 co-cultured with U937 cells, the percentage of cells in S phase (48.860 +/- 2.290), the DNA synthesis [(5694 +/- 917) min(-1)], and the expression of HOXB2 mRNA (0.947 +/- 0.003) were obviously higher than those in control group [41.590 +/- 2.590 vs (2498 +/- 1109) min(-1) vs 0.646 +/- 0.004, P > 0.01]. There was no obvious difference in apoptosis among above stimulation methods (P >0.05).

CONCLUSIONU937 cells activated by ConA can promote the proliferation of ECV304 cells and further regulate angiogenesis. HOXB2 gene is closely related to the endothelial proliferation.

Apoptosis ; Cell Division ; Cell Line ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Endothelium, Vascular ; cytology ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Macrophages ; cytology ; metabolism ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism ; Umbilical Veins ; cytology