cbil.upenn.edu:RAD.Protocol:5897:Protocol - cbil.upenn.edu:RAD.Protocol:5897:Protocol

Type
dissect
Description
Arterial segments were dissected from the surrounding tissue, flushed with cold sterile phosphate-buffered saline (PBS), and cut open lengthwise. Approximately 1,000 to 10,000 endothelial cells were gently scraped from the following regions: (i) Proximal Left Anterior Descending Coronary Artery (LCP): 1 cm-long athero-susceptible region 0.5 cm distal to left coronary bifurcation; (ii) Distal Left Anterior Descending Coronary Artery (LCFD): 1 cm-long athero-protected region 5 cm distal to the left coronary bifurcation; (iii) Proximal Right Coronary Artery (RCP): 1 cm-long athero-susceptible region 0.5 cm distal to the origin of the artery; (iv) Distal Right Coronary Artery (RCFD): 1cm-long athero-protected region 5 cm distal to the origin of the artery; (v) Proximal Left Circumflex Artery (CP): 1 cm-long athero-susceptible region from the origin artery; (vi) Distal Left Circumflex Artery (CD): 1 cm-long athero-protected region 5 cm distal to the origin of the artery; (vii) Aortic Arch (AA): 1-cm^2 athero-susceptible region located at the inner-curve and lateral walls of the aortic arch; (viii) Descending Thoracic Aorta (DT): 1-cm^2 athero-protected region region located proximal to the first intercostal ostia extending to the second intercostal branches in the dorsal descending thoracic aorta; (ix) Brachiocephalic Artery (BA): 1-cm^2 athero-susceptible region at the origin of the artery; (x) Renal Branch (RB): 0.5 cm of proximal-most athero-susceptible region of the renal artery; (xi) Renal Artery (RA): 1 cm-long athero-protected region 1.5 cm distal to the renal artery origin; (xii) Abdominal Aorta (AbA): 1-cm^2 athero-susceptible region 2 cm distal to the renal arteries; (xiii) Common Carotid Artery (C): 1 cm^2 athero-protected region in the mid-section of the artery. The cells were transferred directly to lysis buffer containing RNase inhibitors guanidine isothiocyanate and beta-mercaptoethanol (0.143 M) (Stratagene, La Jolla, CA) and stored on dry ice.
Links
All experiments using protocol cbil.upenn.edu:RAD.Protocol:5897:Protocol: (E-CBIL-42, E-CBIL-43)