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Preparation and characterization of human gingival cells
Authors:Eleana D Stoufi  Martin A Taubman  Jeffrey L Ebersole  Daniel J Smith
Institution:Department of Immunology, Forsyth Dental Center;Department of Oral Pathology, Harvard School of Dental Medicine, Boston, MA, U.S.A.
Abstract:A procedure was developed for the preparation of single cell suspensions from gingival and periodontal tissues using collagenase digestion followed by gentle mechanical disruption. The procedure was evaluated on rat gingival tissues. One-hour incubation was found to produce the greatest number of cells with the highest viability, and the largest recovery of mononuclear cells. Collagenase did not appear to effect recovery, viability or B cell surface markers on human peripheral blood cells. Human gingival and periodontal tissues were obtained from 35 patients: 20 with adult periodontitis; 8 with juvenile periodontitis; 7 with normal gingivae or chronic gingivitis. Scaling, followed by probing measurements and then surgery of the affected sites was routinely performed. In single cell suspensions from tissue obtained from surgery, significantly fewer mononuclear cells were recovered from the tissues of normal/chronic gingivitis patients than cells/mg of tissue recovered from adult periodontitis or juvenile periodontitis groups. The maximum contribution of blood mononuclear cells to the gingival cells averaged 0.5+0.2%. This method is useful for recovering gingival (periodontal) cells for phenotypic and functional studies.
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