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Optimized medium for clonal growth of human microvascular endothelial cells with minimal serum

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Summary

An optimized basal nutrient medium, MCBD 131, has been developed that supports clonal growth of human microvascular endothelial cells (HMVEC) with as little as 0.7% dialyzed fetal bovine serum (dFBS) when also supplemented with 10 ng/ml epidermal growth factor (EGF) and 1 μg/ml hydrocortisone. An extensive initial survey of available media showed that MCDB 402, a medium optimized for low-serum growth of Swiss 3T3 cells, supported the best clonal growth of HMVEC with 10% dFBS. Quantitative adjustment of the composition of MCDB 402 for improved clonal growth of HMVEC with reduced amounts of dFBS resulted in development of MCDB 131. Although many different adjustments contributed to the optimal properties of MCDB 131 for growth of HMVEC, the most unusual feature of this medium is its high magnesium concentration. A major benefit was achieved by increasing Mg2+ from 0.8 mM in MCDB 402 to 10.0 mM in MCDB 131. In the absence of defined supplements, MCDB 131 supports good clonal growth of HMVEC with 2% dFBS. This can be reduced to 0.7% by adding EGF and hydrocortisone, which act synergistically to improve growth with low levels of dFBS.

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  1. Ann Knedler

    Present address: Department of Pediatrics, National Jewish Center for Immunology and Respiratory Medicine, 1400 Jackson, 80206, Denver, Colorado

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  1. Department of Molecular, Cellular and Developmental Biology, University of Colorado, Campus Box 347, 80309, Boulder, Colorado

    Ann Knedler & Richard G. Ham

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This research was supported by grant CA 15305 from the National Cancer Institute, Bethesda, MD.

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Knedler, A., Ham, R.G. Optimized medium for clonal growth of human microvascular endothelial cells with minimal serum. In Vitro Cell Dev Biol 23, 481–491 (1987). https://doi.org/10.1007/BF02628418

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  • DOI: https://doi.org/10.1007/BF02628418

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