Organ culture of C57BL/6 mouse arteries with LPS as an in vitro model of vascular inflammation

TY - CONF

T1 - Organ culture of C57BL/6 mouse arteries with LPS as an in vitro model of vascular inflammation

AU - Outzen, Emilie Middelbo

AU - Mehryar, Rahila

AU - Boonen, Harrie C.M.

AU - Nielsen, Anette Sams

AU - Sheykhzade, Majid

PY - 2013/8/28

Y1 - 2013/8/28

N2 - Background: Vascular inflammation is believed to be involved in the pathogenesis of various cardiovascular diseases, the study of which often involves use of the mouse strain C57BL/6. In vivo studies can, however, be difficult to control and interpret. Aim of the study: To set up and characterise an in vitro model for studying vascular inflammation and function in cultured arteries from C57BL/6 mice. Methods: Segments of abdominal aorta and mesenteric arteries (MA) were incubated for 24 hours at 37̊C and 95% O2/5% CO2 in DMEM ± 100 ng/mL LPS. Aorta segments were frozen for molecular studies of inflammatory markers. MA segments were mounted in tissue baths for measurement of isometric contractile force. Functional studies of MA included characterisation of length-tension relationship and pharmacological reactivity to selected agonists.Results and discussion: Maximum active wall tension of C57BL/6 MA was achieved at a normalisation factor of 0.9 (0.91 ± 0.06, mean ± SEM, n = 9) as observed (0.85 ± 0.06, mean ± SEM, n = 3) and previously described in rat MA (Mulvany and Halpern, 1977). Furthermore, preliminary findings show that organ culture with 100 ng/mL LPS decreases endothelium-dependent dilation of C57BL/6 MA along with increased expression of inflammatory markers in aorta. Conclusions: In C57BL/6 MA, maximum active wall tension was achieved with a normalisation factor of 0.9. Furthermore, organ culture with LPS induces vascular inflammation and functional changes in C57BL/6 arteries. The in vitro model allows further assessment of relevant vasoactive systems and studies of possible protective mediators.

AB - Background: Vascular inflammation is believed to be involved in the pathogenesis of various cardiovascular diseases, the study of which often involves use of the mouse strain C57BL/6. In vivo studies can, however, be difficult to control and interpret. Aim of the study: To set up and characterise an in vitro model for studying vascular inflammation and function in cultured arteries from C57BL/6 mice. Methods: Segments of abdominal aorta and mesenteric arteries (MA) were incubated for 24 hours at 37̊C and 95% O2/5% CO2 in DMEM ± 100 ng/mL LPS. Aorta segments were frozen for molecular studies of inflammatory markers. MA segments were mounted in tissue baths for measurement of isometric contractile force. Functional studies of MA included characterisation of length-tension relationship and pharmacological reactivity to selected agonists.Results and discussion: Maximum active wall tension of C57BL/6 MA was achieved at a normalisation factor of 0.9 (0.91 ± 0.06, mean ± SEM, n = 9) as observed (0.85 ± 0.06, mean ± SEM, n = 3) and previously described in rat MA (Mulvany and Halpern, 1977). Furthermore, preliminary findings show that organ culture with 100 ng/mL LPS decreases endothelium-dependent dilation of C57BL/6 MA along with increased expression of inflammatory markers in aorta. Conclusions: In C57BL/6 MA, maximum active wall tension was achieved with a normalisation factor of 0.9. Furthermore, organ culture with LPS induces vascular inflammation and functional changes in C57BL/6 arteries. The in vitro model allows further assessment of relevant vasoactive systems and studies of possible protective mediators.

KW - Former Faculty of Pharmaceutical Sciences

KW - Vascular

KW - Inflammation

KW - organ culture

KW - Lipopolysaccharides

KW - Adhesion factors

M3 - Poster

T2 - DRA Summer School 2013

Y2 - 28 August 2013 through 29 August 2013

ER -