Abstract
The pericellular matrix of human fibroblast cultures was isolated, using sequential extraction with sodium deoxycholate and hypotonic buffer in the presence of protease inhibitor. The matrix attached to the growth substratum had a "sackcloth-like" structure as seen by phase contrast, immunofluorescence, and scanning electron microscopy, and it had a vaguely filamentous ultrastructure similar to that seen in intact cell layers. The matrix consisted of hyaluronic acid and heparan sulfate as the major glycosaminoglycan components and fibronectin and procollagen as major polypeptides as shown by metabolic labeling, gel electrophoresis, immunofluorescence, and collagenase digestion. This pericellular matrix can be regarded as an in vitro equivalent of the loose connective tissue matrix.
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- Culp L. A. Electrophoretic analysis of substrate-attached proteins from normal and virus-transformed cells. Biochemistry. 1976 Sep 7;15(18):4094–4104. doi: 10.1021/bi00663a028. [DOI] [PubMed] [Google Scholar]
- Hascall V. C., Heinegård D. Aggregation of cartilage proteoglycans. II. Oligosaccharide competitors of the proteoglycan-hyaluronic acid interaction. J Biol Chem. 1974 Jul 10;249(13):4242–4249. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
- Olden K., Yamada K. M. Direct detection of antigens in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1977 Apr;78(2):483–490. doi: 10.1016/0003-2697(77)90108-7. [DOI] [PubMed] [Google Scholar]
- Rice R. H., Means G. E. Radioactive labeling of proteins in vitro. J Biol Chem. 1971 Feb 10;246(3):831–832. [PubMed] [Google Scholar]
- Rosen J. J., Culp L. A. Morphology and cellular origins of substrate-attached material from mouse fibroblasts. Exp Cell Res. 1977 Jun;107(1):139–149. doi: 10.1016/0014-4827(77)90395-0. [DOI] [PubMed] [Google Scholar]