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Isolation, growth requirements, cloning, prostacyclin production and life-span of human adult endothelial cells in low serum culture medium

  • Rapid Communications in Cell Biology
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Editor's statement This paper provides an opportunity for relatively rapid, easy growth and cloning of endothelial cells from various human specimens which are more difficult to deal with than those obtained from an intact artery or intact umbilical vein. Russel Ross

Summary

Endothelial cells from autopsy and biopsy specimens from a variety of adult human vascular tissue were harvested by collagenase treatment and gentle swabbing of the lumenal surface. Nutrient medium MCDB 107 containing a partially purified brain-derived growth factor (5 μg/ml), epidermal growth factor (10 ng/ml) and only 2% (v/v) fetal bovine serum supported clonal and long-term serial culture (17.6 to 26.1 cumulative population doublings) of endothelial cells from vena cava, thoracic aorta and tibial arteries at a 70% rate of success. Cumulative doublings of the cell population from eight cultures were inversely proportional to age of donor of the vascular tissue from which cells were isolated. Heparin had an enhancing effect on cell growth that varied with cell strain. Prostacyclin production of human adult endothelial cell cultures was stimulated by aracidonate and thrombin by 17 to 20 and 2 to 3-fold respectively. Endogenous and stimulated rates of prostacyclin production by human adult endothelial cells were 2 to 3 times that of human adult smooth muscle cells and 20 to 30 times that of human fibroblasts.

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Authors and Affiliations

  1. W. Alton Jones Cell Science Center, Inc., 10 Old Barn Road, 12946, Lake Placid, New York

    Hiroyoshi Hoshi & Wallace L. McKeehan

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  1. Hiroyoshi Hoshi
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  2. Wallace L. McKeehan
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The work was supported by Public Health Service Grant AGO3275 and Grant No. 1718 from the Council for Tobacco Research.

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Hoshi, H., McKeehan, W.L. Isolation, growth requirements, cloning, prostacyclin production and life-span of human adult endothelial cells in low serum culture medium. In Vitro Cell Dev Biol 22, 51–56 (1986). https://doi.org/10.1007/BF02623441

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  • DOI: https://doi.org/10.1007/BF02623441

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