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. 1970 Oct 1;132(4):765–774. doi: 10.1084/jem.132.4.765

FURTHER STUDIES ON METACHROMASIA IN CULTURED HUMAN FIBROBLASTS

STAINING OF GLYCOSAMINOGLYCANS (MUCOPOLYSACCHARIDES) BY ALCIAN BLUE IN SALT SOLUTIONS

B Shannon Danes 1, J E Scott 1, Alexander G Bearn 1
PMCID: PMC2138861  PMID: 4101365

Abstract

Staining with Alcian blue in various concentrations of magnesium chloride (alcianophilia) has been found to be a useful supplement to metachromatic staining to detect increased cellular concentrations of glycosaminoglycans (mucopolysaccharides). In many instances alcianophilia at 0.3 M MgCl2 is more specific than metachromasia and does not give "false positives" sometimes found in normal individuals and in those with cystic fibrosis, Gaucher’s disease, familial amaurotic idiocy, and pseudoxanthoma elasticum. On the other hand, it gives a "false negative" reaction in the Sanfilippo syndrome (perhaps because the characteristically elevated glycosaminoglycan in this disease, heparan sulfate, is not synthesized by cultured skin fibroblasts), and in the Marfan syndrome. It detects the Maroteaux-Lamy syndrome, which metachromasia does not. The "false positives" given by metachromasia in all six families studied thus far are genuine, reproducible reactions that can be traced through at least three generations of normal individuals within a family. There is therefore, in these families, a genetic factor that causes such metachromasia, but it is not increased glycosaminoglycan concentration.

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Selected References

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

  1. Bearn A. G., Danes B. S. A genetic study of cystic fibrosis of the pancreas in cell culture. Trans Assoc Am Physicians. 1969;82:248–252. [PubMed] [Google Scholar]
  2. Bearn A. G., Danes B. S. Metachromasia elaborated. N Engl J Med. 1970 Jan 8;282(2):102–102. doi: 10.1056/NEJM197001082820217. [DOI] [PubMed] [Google Scholar]
  3. Danes B. S., Bearn A. G. Cystic fibrosis of the pancreas. A study in cell culture. J Exp Med. 1969 Apr 1;129(4):775–793. doi: 10.1084/jem.129.4.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Danes B. S., Bearn A. G. Cystic fibrosis: distribution of mucopolysaccharides in fibroblast cultures. Biochem Biophys Res Commun. 1969 Sep 10;36(6):919–924. doi: 10.1016/0006-291x(69)90291-5. [DOI] [PubMed] [Google Scholar]
  5. Danes B. S., Bearn A. G. Hurler's syndrome. A genetic study in cell culture. J Exp Med. 1966 Jan 1;123(1):1–16. doi: 10.1084/jem.123.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Matalon R., Dorfman A. Acid mucopolysaccharides in cultured fibroblasts of cystic fibrosis of the pancreas. Biochem Biophys Res Commun. 1968 Dec 30;33(6):954–958. doi: 10.1016/0006-291x(68)90405-1. [DOI] [PubMed] [Google Scholar]
  7. Matalon R., Dorfman A. Acid mucopolysaccharides in cultured human fibroblasts. Lancet. 1969 Oct 18;2(7625):838–841. doi: 10.1016/s0140-6736(69)92289-2. [DOI] [PubMed] [Google Scholar]
  8. Matalon R., Dorfman A. Hurler's syndrome: biosynthesis of acid mucopolysaccharides in tissue culture. Proc Natl Acad Sci U S A. 1966 Oct;56(4):1310–1316. doi: 10.1073/pnas.56.4.1310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Matalon R., Dorfman A. The accumulation of hyaluronic acid in cultured fibroblasts of the Marfan syndrome. Biochem Biophys Res Commun. 1968 Jul 26;32(2):150–154. doi: 10.1016/0006-291x(68)90361-6. [DOI] [PubMed] [Google Scholar]
  10. Meyer K. Biochemistry and biology of mucopolysaccharides. Am J Med. 1969 Nov;47(5):664–672. doi: 10.1016/0002-9343(69)90162-4. [DOI] [PubMed] [Google Scholar]
  11. Milunsky A., Littlefield J. W. Diagnostic limitations of metachromasia. N Engl J Med. 1969 Nov 13;281(20):1128–1129. doi: 10.1056/NEJM196911132812013. [DOI] [PubMed] [Google Scholar]
  12. Nadler H. L., Wodnicki J. M., Swae M. A., O'Flynn M. E. Cultivated amniotic-fluid cells and fibroblasts derived from families with cystic fibrosis. Lancet. 1969 Jul 12;2(7611):84–85. doi: 10.1016/s0140-6736(69)92393-9. [DOI] [PubMed] [Google Scholar]
  13. SCOTT J. E. Aliphatic ammonium salts in the assay of acidic polysaccharides from tissues. Methods Biochem Anal. 1960;8:145–197. doi: 10.1002/9780470110249.ch4. [DOI] [PubMed] [Google Scholar]
  14. Scott J. E., Dorling J. Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemie. 1965 Oct 1;5(3):221–233. doi: 10.1007/BF00306130. [DOI] [PubMed] [Google Scholar]
  15. Taysi K., Kistenmacher M. L., Punnett H. H., Mellman W. J. Limitations of metachromasia as a diagnostic aid in pediatrics. N Engl J Med. 1969 Nov 13;281(20):1108–1111. doi: 10.1056/NEJM196911132812009. [DOI] [PubMed] [Google Scholar]

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