Protocol for Aortic Ring Assay
来自 : 蚂蚁淘
Procedures
- Covera48-wellplatewithMatrigel(100μl/well)ofandincubatefor30minat37°C,5%CO2.
- Sacrificethe1-2montholdmice/rats(WT/mutant)andexcisethoracicaorta.Transfertheaortaintoadishwithsterilized1xPBSbuffer.AfterremovingthefibroADIposetissue,sectionarteriesinto1-1.5mmlongcrosssections,rinsethesectionsfivetimeswithendothelialcellgrowthmedium(ECGM),andplacethemontheMatrigel-coatedwells.
- Coverthearteryringswithanadditional100μlofMatrigel.Iftherearesomebubbles,usePipettetoremovethem.Incubatetheringsin1.5mlofECGMmedium.
- After24hourincubation,replacethemediumwith1.5mlofendothelialcellgrowthmedium(orcontainingwiththeinterestingpeptides/compounds/others).
- After2days,themediumwasreplacedwithmediumoftheexactcompositionasdescribedinstep4.
- Attheday4,takeimagesofaorticrings.Delineatetheoutgrowthareaandmeasuretheoutgrowthmicro-vesselwiththeImageProPlussoftware(MediaCybernetic).
免责声明 本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容。
版权声明 未经蚂蚁淘授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘”。违反上述声明者，本网将追究其相关法律责任。
