Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1979 Apr;76(4):1809–1813. doi: 10.1073/pnas.76.4.1809

Purification of human platelet-derived growth factor.

H N Antoniades, C D Scher, C D Stiles
PMCID: PMC383481  PMID: 287022

Abstract

Human platelets contain a polypeptide growth factor that stimulates the proliferation of connective tissue cells. Purification of this platelet-derived growth factor (PDGF) was accomplished by heat (100 degrees C) treatment of washed platelets and subsequent ion-exchange chromatography, gel filtration in 1 M acetic acid, isoelectric focusing, and preparative sodium dodecyl sulfate/polyacrylamide gel electrophoresis. PDGF has an isoelectric point of 9.8 and a molecular weight ranging from 13,000 to 16,000 as judged by gel filtration in 1 M acetic acid or analytical sodium dodecyl sulfate gel electrophoresis under reducing conditions. The specific activity of the purified PDGF is 20 million times greater than that found in unfractionated human serum. Purified PDGF stimulates replicative DNA synthesis and cell proliferation in quiescent density-arrested cultures of BALB/c 3T3 cells at concentrations of 1 ng/ml (0.1 nM).

Full text

PDF
1809

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Antoniades H. N., Scher C. D. Growth factors derived from human serum, platelets, and pituitary: properties and immunologic cross-reactivity. Natl Cancer Inst Monogr. 1978 May;(48):137–140. [PubMed] [Google Scholar]
  2. Antoniades H. N., Scher C. D. Radioimmunoassay of a human serum growth factor for Balb/c-3T3 cells: derivation from platelets. Proc Natl Acad Sci U S A. 1977 May;74(5):1973–1977. doi: 10.1073/pnas.74.5.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Armelin H. A. Pituitary extracts and steroid hormones in the control of 3T3 cell growth. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2702–2706. doi: 10.1073/pnas.70.9.2702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. COHEN S. Isolation of a mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the new-born animal. J Biol Chem. 1962 May;237:1555–1562. [PubMed] [Google Scholar]
  5. Dulbecco R. Topoinhibition and serum requirement of transformed and untransformed cells. Nature. 1970 Aug 22;227(5260):802–806. doi: 10.1038/227802a0. [DOI] [PubMed] [Google Scholar]
  6. Froesch E. R., Zapf J., Audhya T. K., Ben-Porath E., Segen B. J., Gibson K. D. Nonsuppressible insulin-like activity and thyroid hormones: major pituitary-dependent sulfation factors for chick embryo cartilage. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2904–2908. doi: 10.1073/pnas.73.8.2904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gospodarowicz D. Localisation of a fibroblast growth factor and its effect alone and with hydrocortisone on 3T3 cell growth. Nature. 1974 May 10;249(453):123–127. doi: 10.1038/249123a0. [DOI] [PubMed] [Google Scholar]
  8. Hayashi I., Sato G. H. Replacement of serum by hormones permits growth of cells in a defined medium. Nature. 1976 Jan 15;259(5539):132–134. doi: 10.1038/259132a0. [DOI] [PubMed] [Google Scholar]
  9. Heldin C. H., Wasteson A., Westermark B. Partial purification and characterization of platelet factors stimulating the multiplication of normal human glial cells. Exp Cell Res. 1977 Oct 15;109(2):429–437. doi: 10.1016/0014-4827(77)90023-4. [DOI] [PubMed] [Google Scholar]
  10. Holley R. W. Control of growth of mammalian cells in cell culture. Nature. 1975 Dec 11;258(5535):487–490. doi: 10.1038/258487a0. [DOI] [PubMed] [Google Scholar]
  11. Holley R. W., Kiernan J. A. "Contact inhibition" of cell division in 3T3 cells. Proc Natl Acad Sci U S A. 1968 May;60(1):300–304. doi: 10.1073/pnas.60.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kohler N., Lipton A. Platelets as a source of fibroblast growth-promoting activity. Exp Cell Res. 1974 Aug;87(2):297–301. doi: 10.1016/0014-4827(74)90484-4. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Pierson R. W., Jr, Temin H. M. The partial purification from calf serum of a fraction with multiplication-stimulating activity for chicken fibroblasts in cell culture and with non-suppressible insulin-like activity. J Cell Physiol. 1972 Jun;79(3):319–330. doi: 10.1002/jcp.1040790302. [DOI] [PubMed] [Google Scholar]
  15. Pledger W. J., Stiles C. D., Antoniades H. N., Scher C. D. Induction of DNA synthesis in BALB/c 3T3 cells by serum components: reevaluation of the commitment process. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4481–4485. doi: 10.1073/pnas.74.10.4481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ross R., Glomset J., Kariya B., Harker L. A platelet-dependent serum factor that stimulates the proliferation of arterial smooth muscle cells in vitro. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1207–1210. doi: 10.1073/pnas.71.4.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ross R., Glomset J., Kariya B., Raines E. Role of platelet factors in the growth of cells in culture. Natl Cancer Inst Monogr. 1978 May;(48):103–108. [PubMed] [Google Scholar]
  18. Rubin H., Koide T. Stimulation of DNA synthesis and 2-deoxy-D-glucose transport in chick embryo cultures by excessive metal concentrations and by a carcinogenic hydrocarbon. J Cell Physiol. 1973 Jun;81(3):387–396. doi: 10.1002/jcp.1040810311. [DOI] [PubMed] [Google Scholar]
  19. Scher C. D., Stathakos D., Antoniades H. N. Dissociation of cell division stimulating capacity for Balb-c-3T3 from the insulin-like activity in human serum. Nature. 1974 Feb 1;247(5439):279–281. doi: 10.1038/247279a0. [DOI] [PubMed] [Google Scholar]
  20. Temin H. M. Studies on carcinogenesis by avian sarcoma viruses. 3. The differential effect of serum and polyanions on multiplication of uninfected and converted cells. J Natl Cancer Inst. 1966 Aug;37(2):167–175. [PubMed] [Google Scholar]
  21. Todaro G. J., Lazar G. K., Green H. The initiation of cell division in a contact-inhibited mammalian cell line. J Cell Physiol. 1965 Dec;66(3):325–333. doi: 10.1002/jcp.1030660310. [DOI] [PubMed] [Google Scholar]
  22. Udenfriend S., Stein S., Böhlen P., Dairman W., Leimgruber W., Weigele M. Fluorescamine: a reagent for assay of amino acids, peptides, proteins, and primary amines in the picomole range. Science. 1972 Nov 24;178(4063):871–872. doi: 10.1126/science.178.4063.871. [DOI] [PubMed] [Google Scholar]
  23. Weber K., Kuter D. J. Reversible denaturation of enzymes by sodium dodecyl sulfate. J Biol Chem. 1971 Jul 25;246(14):4504–4509. [PubMed] [Google Scholar]
  24. Wiebel F., Baserga R. Early alterations in amino acid pools and protein synthesis of diploid fibroblasts stimulated to synthesize DNA by addition of serum. J Cell Physiol. 1969 Oct;74(2):191–202. doi: 10.1002/jcp.1040740211. [DOI] [PubMed] [Google Scholar]
  25. van Wyk J. J., Underwood L. E., Baseman J. B., Hintz R. L., Clemmons D. R., Marshall R. N. Explorations of the insulinlike and growth-promoting properties of somatomedin by membrane receptor assays. Adv Metab Disord. 1975;8:127–150. doi: 10.1016/b978-0-12-027308-9.50015-9. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES