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Localization of mucopolysaccharides in epithelial‐Iike cells cultured in vitro
Author(s) -
Thonard J. C.,
Wiebkin O. W.
Publication year - 1973
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/j.1600-0765.1973.tb00749.x
Subject(s) - cetylpyridinium chloride , staining , nucleic acid , toluidine , glycosaminoglycan , biochemistry , in vitro , chemistry , tissue culture , microbiology and biotechnology , ethanol precipitation , sudan black b , biology , polysaccharide , genetics , pulmonary surfactant
Previous studies, demonstrating mucopolysaccharides (MPS) in tissue cultured epithelial cells did not eliminate the possibility that some of the reported staining could be attributed to acidic lipids, nucleic acids and substrate movement, although these possibilities were deemed minimal due to the employment of generally accepted techniques. Human amnion cells were grown on coverslips in tissue culture. Some coverslips were treated to remove MPS, lipid or nucleic acids before fixation and staining, whilst others were merely fixed and stained. Fixatives included 10 % formalin. 10 % formalin in 80 % ethanol, 1 % cetylpyridinium chloride and 1 % cetylpyridinium chloride in 80 % ethanol. Stains employed were the Hale colloidal iron, alcian blue, toluidine blue at pH 6.0 and 2.5 and sudan black. Substrate extractions were effected with testicular hyaluronidase, ribonuclease, KOH and pyridirje. The present, report confirms earlier results and provides evidence that human epithelial‐like cells produce MPS in vitro the staining specificity of which eliminates lipids and nucleic acids as the source of the observed reactions. It is claimed that the MPS is mainly confined to the cell surface and therefore is more discernible as a MPS stainable moiety in contiguous cell cultures than when cells are dispersed singly.