The Effect of Human Angiopoietin-1 on the Proliferation and Apoptosis of Human Umbilical Vein Endothelial Cell
WU Kai-chun
Abstract
WU Kai-chun
Abstract
Objective: To investigate the effect of human Angiopoietin-1 (Ang-1) on the proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC). Methods: The recombinant eukaryotic expression vector pcDNA3. 1-V5-HisC-Angl was constructed and was used to transiently transfecte to 293 cells. HUVEC was seperated from new born fetel cord. The supernatants from transfected 293 cells were harvested, and then added to HUVEC culture. The proliferation and apoptosis of HUVEC was measured by MTT colorimetry assay analysis, cell counting and flow cytometry, respectively. Results: Proliferation of HUVEC cells cultured with supurnatants of Ang-1 transfected group was higher than those untreated and mock transfected group, results of which were 0. 68 ±0. 10, 0. 36 ± 0. 11, 0. 40 ± 0. 03 in the MTT assays; and were ( 15. 03 ± 1. 98) × 104 , ( 10. 13 ±2. 06) × 104 and (8. 7 ± 1. 73) × 104 respectively in the cell counts. The ap-optotic pereentage of HUVEC cells culturel with supernatants from Ang-1 transfected group was lower than those of untreated or mock transfeeted group, results of which were 6% ,21% and 19% respectively in FACS analysis. Conclusions: Human Ang-1 can significacetly pronaete stimulate the proliferation and reduce the apoptosis of endothelial cells.
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Objective: To investigate the effect of human Angiopoietin-1 (Ang-1) on the proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC). Methods: The recombinant eukaryotic expression vector pcDNA3. 1-V5-HisC-Angl was constructed and was used to transiently transfecte to 293 cells. HUVEC was seperated from new born fetel cord. The supernatants from transfected 293 cells were harvested, and then added to HUVEC culture. The proliferation and apoptosis of HUVEC was measured by MTT colorimetry assay analysis, cell counting and flow cytometry, respectively. Results: Proliferation of HUVEC cells cultured with supurnatants of Ang-1 transfected group was higher than those untreated and mock transfected group, results of which were 0. 68 ±0. 10, 0. 36 ± 0. 11, 0. 40 ± 0. 03 in the MTT assays; and were ( 15. 03 ± 1. 98) × 104 , ( 10. 13 ±2. 06) × 104 and (8. 7 ± 1. 73) × 104 respectively in the cell counts. The ap-optotic pereentage of HUVEC cells culturel with supernatants from Ang-1 transfected group was lower than those of untreated or mock transfeeted group, results of which were 6% ,21% and 19% respectively in FACS analysis. Conclusions: Human Ang-1 can significacetly pronaete stimulate the proliferation and reduce the apoptosis of endothelial cells.
Key concepts: Umbilical vein, Apoptosis, Transfection, Flow cytometry, Human umbilical vein endothelial cell, Molecular biology, Cell growth, MTT assay