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Clonal growth of differentiated rabbit cartilage cells
Author(s) -
Ham Richard G.,
Sattler Gerald L.
Publication year - 1968
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040720205
Subject(s) - metachromasia , staining , cartilage , collagenase , phase contrast microscopy , fetal bovine serum , incubation period , biology , matrix (chemical analysis) , incubation , anatomy , cell , andrology , pathology , chemistry , microbiology and biotechnology , biochemistry , medicine , enzyme , chromatography , physics , optics
Abstract Single cell suspensions have been prepared from rabbit cartilage by collagenase digestion in the presence of fetal bovine serum. When inoculated into medium F12 supplemented with 5% fetal bovine serum and 5% rabbit serum, the cartilage cell suspensions yield large healthy colonies of epithelial‐like cells with an average plating efficiency of 47%. At the end of a 14 day incubation period, the centers of many of the colonies are thickened into multilayered aggregates. Such colonies exhibit serveral cartilage‐like properties, including metachromatic staining with toluidine blue, dense brown staining with bismark brown, bright green staining with alcian green, and a refractile matrix which can be seen with the phase contrast microscope to surround the living cells. Similar properties have also been observed in second and third passage subcultures.