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. 1984 Oct;81(20):6413–6417. doi: 10.1073/pnas.81.20.6413

Brain- and liver cell-derived factors are required for growth of human endothelial cells in serum-free culture.

H Hoshi, W L McKeehan
PMCID: PMC391934  PMID: 6333682

Abstract

A test of the mitogenic activity of greater than 40 purified and crude sources of hormones and growth factors revealed that epidermal growth factor, high density lipoprotein, an extract of bovine pituitary, hypothalamus, or whole brain, and the medium conditioned by differentiated human hepatoma cells were mitogenic for cultured endothelial cells derived from human umbilical vein. The four active agents combined with an improved nutrient medium and a collagen- or fibronectin-coated culture surface supported the growth of the endothelial cell population at a rate of 0.70-0.80 generations per day at both low and high cell densities in serum-free medium. The brain-derived activity exhibited properties reported by Maciag et al. [Maciag, T., Hoover, A. & Weinstein, R. (1982) J. Biol. Chem. 257, 5333-5336] and Gordon et al. [Gordon, P. B., Sussman, I. I. & Hatcher, V. B. (1983) In Vitro 19, 661-671]. The liver cell-derived activity was a specific product of differentiated hepatoma cells. The medium from HeLa cells, relatively undifferentiated rat liver cell lines, and human fibroblasts was inactive. Purified plasma proteins of liver origin could not substitute for the hepatoma cell-conditioned medium. The hepatoma cell-derived activity was non-dialyzable, heat-labile, stable between pH 4 to 11, inactivated by trypsin and mercaptoethanol treatment, and stable after treatment with 6 M urea and phenylmethylsulfonyl fluoride. The results provide a simplified model for elucidation of the endocrinology of human endothelial cell growth, function, and aging. We suggest an endocrine role of both the nervous system and liver in the regeneration of endothelial cells.

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Selected References

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  1. Alexander J. J., Bey E. M., Geddes E. W., Lecatsas G. Establishment of a continuously growing cell line from primary carcinoma of the liver. S Afr Med J. 1976 Dec 18;50(54):2124–2128. [PubMed] [Google Scholar]
  2. Barnes D. W., Silnutzer J. Isolation of human serum spreading factor. J Biol Chem. 1983 Oct 25;258(20):12548–12552. [PubMed] [Google Scholar]
  3. Barnes D., Sato G. Methods for growth of cultured cells in serum-free medium. Anal Biochem. 1980 Mar 1;102(2):255–270. doi: 10.1016/0003-2697(80)90151-7. [DOI] [PubMed] [Google Scholar]
  4. Clemmons D. R., Isley W. L., Brown M. T. Dialyzable factor in human serum of platelet origin stimulates endothelial cell replication and growth. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1641–1645. doi: 10.1073/pnas.80.6.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gordon P. B., Sussman I. I., Hatcher V. B. Long-term culture of human endothelial cells. In Vitro. 1983 Sep;19(9):661–671. doi: 10.1007/BF02628957. [DOI] [PubMed] [Google Scholar]
  6. Gospodarowicz D., Brown K. D., Birdwell C. R., Zetter B. R. Control of proliferation of human vascular endothelial cells. Characterization of the response of human umbilical vein endothelial cells to fibroblast growth factor, epidermal growth factor, and thrombin. J Cell Biol. 1978 Jun;77(3):774–788. doi: 10.1083/jcb.77.3.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gospodarowicz D., Cheng J., Lirette M. Bovine brain and pituitary fibroblast growth factors: comparison of their abilities to support the proliferation of human and bovine vascular endothelial cells. J Cell Biol. 1983 Dec;97(6):1677–1685. doi: 10.1083/jcb.97.6.1677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ham R. G., McKeehan W. L. Media and growth requirements. Methods Enzymol. 1979;58:44–93. doi: 10.1016/s0076-6879(79)58126-9. [DOI] [PubMed] [Google Scholar]
  9. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Knauer D. J., Cunningham D. D. A reevaluation of the response of human umbilical vein endothelial cells to certain growth factors. J Cell Physiol. 1983 Dec;117(3):397–406. doi: 10.1002/jcp.1041170315. [DOI] [PubMed] [Google Scholar]
  11. Knowles B. B., Howe C. C., Aden D. P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science. 1980 Jul 25;209(4455):497–499. doi: 10.1126/science.6248960. [DOI] [PubMed] [Google Scholar]
  12. Lemmon S. K., Riley M. C., Thomas K. A., Hoover G. A., Maciag T., Bradshaw R. A. Bovine fibroblast growth factor: comparison of brain and pituitary preparations. J Cell Biol. 1982 Oct;95(1):162–169. doi: 10.1083/jcb.95.1.162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Maciag T., Cerundolo J., Ilsley S., Kelley P. R., Forand R. An endothelial cell growth factor from bovine hypothalamus: identification and partial characterization. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5674–5678. doi: 10.1073/pnas.76.11.5674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maciag T., Hoover G. A., Weinstein R. High and low molecular weight forms of endothelial cell growth factor. J Biol Chem. 1982 May 25;257(10):5333–5336. [PubMed] [Google Scholar]
  15. McKeehan W. L., Adams P. S., Rosser M. P. Modified nutrient medium MCDB 151, defined growth factors, cholera toxin, pituitary factors, and horse serum support epithelial cell and suppress fibroblast proliferation in primary cultures of rat ventral prostate cells. In Vitro. 1982 Feb;18(2):87–91. doi: 10.1007/BF02796399. [DOI] [PubMed] [Google Scholar]
  16. McKeehan W. L., McKeehan K. A. Calcium, magnesium, and serum factors in multiplication of normal and transformed human lung fibroblasts. In Vitro. 1980 Jun;16(6):475–485. doi: 10.1007/BF02626460. [DOI] [PubMed] [Google Scholar]
  17. Nakabayashi H., Taketa K., Miyano K., Yamane T., Sato J. Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. Cancer Res. 1982 Sep;42(9):3858–3863. [PubMed] [Google Scholar]
  18. Tauber J. P., Cheng J., Massoglia S., Gospodarowicz D. High density lipoproteins and the growth of vascular endothelial cells in serum-free medium. In Vitro. 1981 Jun;17(6):519–530. doi: 10.1007/BF02633513. [DOI] [PubMed] [Google Scholar]
  19. Thornton S. C., Mueller S. N., Levine E. M. Human endothelial cells: use of heparin in cloning and long-term serial cultivation. Science. 1983 Nov 11;222(4624):623–625. doi: 10.1126/science.6635659. [DOI] [PubMed] [Google Scholar]
  20. Zetter B. R., Antoniades H. N. Stimulation of human vascular endothelial cell growth by a platelet-derived growth factor and thrombin. J Supramol Struct. 1979;11(3):361–370. doi: 10.1002/jss.400110311. [DOI] [PubMed] [Google Scholar]

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