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Related Experiment Videos

Serum contains a platelet-derived transforming growth factor

C B Childs, J A Proper, R F Tucker

    Proceedings of the National Academy of Sciences of the United States of America
    |September 1, 1982
    PubMed
    Summary

    Fetal bovine serum contains a platelet-derived growth factor that stimulates colony formation in anchorage-dependent cells. This polypeptide requires disulfide bonds and has properties similar to transforming growth factors.

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    Area of Science:

    • Cell Biology
    • Biochemistry
    • Molecular Biology

    Background:

    • Anchorage-dependent cells, such as mouse AKR-2B and rat NRK cells, typically require specific growth factors for proliferation and colony formation.
    • Fetal bovine serum (FBS) is a common supplement in cell culture media, providing a source of various growth factors.
    • Understanding the specific components within FBS that drive cellular responses is crucial for cell culture optimization and research.

    Purpose of the Study:

    • To identify and characterize the colony-stimulating activity present in fetal bovine serum.
    • To determine the origin and biochemical properties of the factor responsible for inducing colony formation.
    • To investigate the relationship between this factor and known growth factors like epidermal growth factor and transforming growth factors.

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    Main Methods:

    • Soft agar colony formation assays using anchorage-dependent mouse AKR-2B and rat NRK cells.
    • Biochemical purification techniques including ammonium sulfate precipitation, molecular sieve chromatography, and isoelectric focusing.
    • Enzymatic and chemical treatments (trypsin, dithiothreitol) to assess protein and disulfide bond requirements.
    • Analysis of platelet-poor plasma and platelet extracts to identify the source of the activity.
    • Comparison of properties with epidermal growth factor receptor binding and transforming growth factors.

    Main Results:

    • Fetal bovine serum induced significant colony formation in soft agar for both cell types.
    • The colony-stimulating activity was characterized as a heat- and acid-stable polypeptide with a molecular weight of 10,000-15,000 Da, requiring disulfide bonds.
    • The activity was found to be of platelet origin, with potent colony-stimulating activity detected in acid/ethanol extracts of platelets.
    • Platelet extracts yielded colony-stimulating activity peaks at 12,000 and 20,000 Da, sharing biochemical properties with the serum activity.
    • No competition for epidermal growth factor receptor binding was observed.

    Conclusions:

    • Serum contains a platelet-derived growth factor(s) that potently stimulates colony formation in anchorage-dependent cells.
    • This platelet-derived factor possesses biochemical properties, including molecular weight and disulfide bond requirement, consistent with a polypeptide.
    • The identified factor exhibits characteristics similar to transforming growth factors, suggesting a potential role in cellular transformation and growth regulation.