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Collagen synthesized and modified by aging fibroblasts in culture

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Summary

Collagen is produced by WI-38 diploid human fibroblast cultures throughout their life cycle. It is examined by a sensitive method based on the analysis of specific peptides obtained after digestion with bacterial collagenase. The production and hydroxylation of the collagen is strongly dependent upon the age (population doublings) of the culture and the presence of ascorbic acid. Young cultures (passage 26) produce large amounts of collagen in the absence of ascorbic acid, and this collagen is about 50% hydroxylated compared to that produced by young cultures in the presence of ascorbic acid. Ascorbic acid reduces to about one-half the amount of collagen produced by these young cultures. The young confluent cultures also depend strongly on ascorbic acid for hydroxylation of proline. The dependence declines rapidly with the age of the culture. The collagen produced by young cultures supplied with ascorbic acid is very similar to the type I collagen produced by normal individuals and has about the same degree of hydroxylation of its prolyl residues. The amount of collagen produced by “older” cultures is unaffected by ascorbic acid, but the degree of hydroxylation is normal only if ascorbic acid is present, and is decreased to about 60 to 70% in the absence of the vitamin. “Senescent” cultures showed little, if any, dependency on ascorbic acid, and the collagen produced, with and without the vitamin, is about 80% hydroxylated.

The prolyl hydroxylation system of the WI-38 cells and the various controls on the system are age-dependent.

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Authors and Affiliations

  1. The Department of Biological Chemistry and Orthopaedic Surgery, Harvard Medical School and Harvard School of Dental Medicine, 300 Longwood Ave., 02115, Boston, Mass.

    Mercedes A. Paz & Paul M. Gallop

  2. The Children's Hospital Medical Center, 300 Longwood Avenue, 02115, Boston, Massachusetts

    Mercedes A. Paz & Paul M. Gallop

  3. Department of Orthopaedic Surgery, Children's Hospital, 300 Longwood Ave., 02115, Boston, Mass.

    Mercedes A. Paz & Paul M. Gallop

Authors
  1. Mercedes A. Paz
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  2. Paul M. Gallop
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Additional information

The work was supported by National Institutes of Health grants AM15671 and HD07376

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Paz, M.A., Gallop, P.M. Collagen synthesized and modified by aging fibroblasts in culture. In Vitro 11, 302–312 (1975). https://doi.org/10.1007/BF02615641

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