Human endothelial cell cultures isolated from internal mammary artery
Abstract
Background: The aim of our study was to prepare homogenic and long term cultures of endothelial cells (EC) from a human internal mammary artery (IMA) obtained during a cardiac by-pass graft operation (CABG). In this way we would be able to assess the conditions suitable for the preparation of sufficient number of autologous arterial ECs and their use in a population mostly affected with cardiovascular diseases.
Methods: Seven 52–76 years old male patients diagnosed with ischemic heart disease were included in our study. Samples of internal mammary artery were collected during CABG performed on these patients. EC cells were prepared from these samples using two approaches: in situ digestion with proteolytical enzymes and proteolytic digestion from the previously resected IMA vessels. Cell morphology and growth kinetics were studied in 3 weeks old cultures. EC cell specificy was determined by immunohystochemical staining for von Willebrand’s factor.
Results: Cell growth could be observed only in cultures prepared with the in situ proteolytic digestion. Cells showed the typical EC morphology and were positive for von Willebrand’s factor. The maximal cell number in colonies (11 in average, 51 at the most) was reached after 6 days in culture. The average cell doubling time of the five cultures was 82 ± 43 hours.
Conclusions: The study proved that our method of isolation is specific for ECs. Using a small IMA sample we were able to collect a number of colonies and sustain them in long term cultures, but the end numbers of cells is still too small for their application in tissue engineering (TE). Our further efforts will be focused on the optimization of the conditions which would allow sufficient proliferation of EC so that small IMA sample would be enough for the TE.
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