[Abstract]  [Full Text]  [PDF]

Silvia Nistri¹, Luca Mazzetti², Paola Failli² and Daniele Bani^1*

¹ Department of Anatomy, Histology and Forensic Medicine, Section of Histology, University of Florence, Italy. V. le G. Pieraccini, 6, I-50139 Florence. Italy. Fax: (+39)055 4271385
² Department of Preclinical and Clinical Pharmacology, University of Florence, Italy. V. le G. Pieraccini, 6, I-50139 Florence. Italy.

* To whom correspondence should be addressed: Daniele Bani, Department of Anatomy, Histology and Forensic Medicine, Section of Histology, University of Florence, Italy. V. le G. Pieraccini, 6, I-50139 Florence. Italy. Fax: (+39)055 4271385. Email: daniele.bani@unifi.it

Biol. Proced. Online 2002;4:32-37. doi:10.1251/bpo31
Submitted: May 30, 2002; Accepted: August 15, 2002; Published: October 28, 2002.

Indexing terms: nitric oxide; Fura-2.

We describe here a method for isolating endothelial cells from rat heart blood vessels by means of coronary microperfusion with collagenase. This methods 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.