High-yield method for isolation and culture of endothelial cells from rat coronary blood vessels suitable for analysis of intracellular calcium and nitric oxide biosynthetic pathwaysReport as inadecuate




High-yield method for isolation and culture of endothelial cells from rat coronary blood vessels suitable for analysis of intracellular calcium and nitric oxide biosynthetic pathways - Download this document for free, or read online. Document in PDF available to download.

Biological Procedures Online

, Volume 4, Issue 1, pp 32–37

Received: 30 May 2002Revised: 14 August 2002Accepted: 15 August 2002DOI: 10.1251-bpo31

Cite this article as: Nistri, S., Mazzetti, L., Failli, P. et al. Biol Proced Online 2002 4: 32. doi:10.1251-bpo31

Abstract

We describe here a method for isolating endothelial cells from rat heart blood vessels by means of coronary microperfusion with collagenase. This method makes it possible to obtain high amounts of endothelial cells in culture which retain the functional properties of their in vivo counterparts, including the ability to uptake fluorescently-labeled acetylated low-density lipoproteins and to respond to vasoactive agents by modulating intracellular calcium and by upregulating intrinsic nitric oxide generation. The main advantages of our technique are: i good reproducibility, ii accurate sterility that can be maintained throughout the isolation procedure and iii high yield of pure endothelial cells, mainly due to microperfusion and temperature-controlled incubation with collagenase which allow an optimal distribution of this enzyme within the coronary vascular bed.

Indexing termsendothelial cells nitric oxide acetylated LDL DAF-2-DA Fura-2AM Published: October 28, 2002

Download to read the full article text



Author: Silvia Nistri - Luca Mazzetti - Paola Failli - Daniele Bani

Source: https://link.springer.com/







Related documents