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Isolation and characterization of human mesenchymal stem cells from gingival connective tissue
Author(s) -
Jin S. H.,
Lee J. E.,
Yun JH.,
Kim I.,
Ko Y.,
Park J. B.
Publication year - 2015
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/jre.12228
Subject(s) - mesenchymal stem cell , connective tissue , stem cell , flow cytometry , microbiology and biotechnology , fibroblast , chemistry , immunofluorescence , cell culture , matrigel , cell , in vitro , biology , pathology , immunology , antibody , biochemistry , medicine , genetics
Background and Objective The main purpose of this study was to isolate and characterize gingival connective tissue‐derived mesenchymal stem cells ( GMSC s). The secondary purpose was to present a modified isolation method for the GMSC s. Material and Methods Collected healthy gingival tissue samples were de‐epithelialized and minced into small fragments. The tissues were digested by dispase and collagenase IV for 30 min. The first digested cell suspension was discarded, and then additional digestion was performed to the remaining cells in the same solution for 90 min. The isolated cells from gingiva was incubated in 37°C humidified condition and observed by inverted microscope. Cytoskeletal morphology was evaluated by phalloidin immunofluorescence. Potency of the cells was tested by colony‐forming unit fibroblast assay. GMSC s were characterized by osteogenic, adipogenic and chondrogenic differentiation, and flow cytometric, immunofluorescence analysis. Results GMSC s showed spindle‐shaped, fibroblast‐like morphology, colony‐forming abilities, adherence to plastic and multilineage differentiation (osteogenic, adipogenic, chondrogenic) potency. GMSC s expressed CD 44, CD 73, CD 90 and CD 105, but did not express CD 14, CD 45, CD 34 and CD 19 in flow cytometry. Expression of stem cell markers ( SSEA ‐4, STRO ‐1, CD 146, CD 166 and CD 271) and a mesenchymal marker (vimentin) were observed by immunofluorescence. Conclusions In conclusion, we isolated and characterized stem cells from human gingival connective tissue with modified protocol. GMSC s showed multipotency with high proliferation and characteristics of mesenchymal stem cells. GMSC s are promising sources for tissue engineering and may be obtained during routine procedures under local anesthesia. Further research is needed to evaluate the potential of GSMC s' proliferation and cryopreservation.