In vitro Study of Proliferation of Human Choroidal Microvascular Endothelial Cells Induced by Vascular Endothelial Growth Factor

Abstract

Abstract: : Purpose: To evaluate the effect of vascular endothelial growth factor(VEGF)on the proliferation of Human Choroidal Microvascular Endothelial Cells (HCMECs) Methods: 1. Using the method of two–step digestion with trypsin and collagenase for human choroidal tissue samples, we obtained a cell suspension. We separated and purified cells by CD31 Dynabeads. HCMECs were then cultured. The HCMECs were identified by morphological evaluation, ultrastructural observation with transmission electron microscopy and immunohistochemical staining of endothelial specific markers, the FVIII–Ag, CD31, and CD34. 2. The HCMECs were divided into these groups: blank, no–serum control group, and no–serum with 100mg/ml VEGF group. Cell proliferation was assayed by MTT at 24hr, 48hr, and 72hr. Results were recorded by optical absorbing value A. 3. Statistical Analysis: The Student’s t test was used for comparison of A values. Statistical software SPSS11 was utilized. Statistical significance was recorded as P<0.05. Results: At 24 hours the mean of A value in non–serum group: 0.096±0.00002,the mean of A value in non–serum with 100mg/ml VEGF group:0.120±0.00003,t=2.45,P<0.05. At 48 hours the mean of A value in non–serum group:0.124±0.0001,the mean of A value in non–serum with 100mg/ml VEGF group:0.163±0.00000,t=2.45,P<0.05. At 72 hours the mean of A value in non–serum group:0.100±0.00005,the mean of A value in non–serum with 100mg/ml VEGF group:0.113±0.00000,t=2.44,P<0.05. Conclusions: VEGF can cause cultured human choroidal vascular endothelial cells to proliferate.

Keywords: vascular cells • proliferation