Isolation and characterization of human mesenchymal stem cells from gingival connective tissue

被引：120
作者：
Jin, S. H. [1 ]
Lee, J. E. [1 ]
Yun, J-H. [2 ]
Kim, I. [3 ]
Ko, Y. [1 ]
Park, J. B. [1 ]
机构：
[1] Catholic Univ Korea, Coll Med, Dept Periodont, Seoul 137701, South Korea
[2] Inha Univ, Sch Med, Div Periodontol, Dept Dent, Inchon, South Korea
[3] Catholic Univ Korea, Coll Med, Uijeongbu St Marys Hosp, Div Oral & Maxillofacial Surg,Dept Dent, Uijongbu, South Korea
来源：
JOURNAL OF PERIODONTAL RESEARCH | 2015年 / 50卷 / 04期
关键词：
gingiva; mesenchymal stromal cells; multipotent stem cells; stem cells; HUMAN ADIPOSE-TISSUE; HUMAN BONE-MARROW; IN-VITRO; PERIODONTAL-LIGAMENT; DENTAL-PULP; COLLAGENASE; EXPANSION; DISPASE; REGENERATION; CULTURES;
D O I：
10.1111/jre.12228
中图分类号：
R78 [口腔科学];
学科分类号：
1003 ;
摘要：
Background and ObjectiveThe main purpose of this study was to isolate and characterize gingival connective tissue-derived mesenchymal stem cells (GMSCs). The secondary purpose was to present a modified isolation method for the GMSCs. Material and MethodsCollected healthy gingival tissue samples were de-epithelialized and minced into small fragments. The tissues were digested by dispase and collagenase IV for 30min. The first digested cell suspension was discarded, and then additional digestion was performed to the remaining cells in the same solution for 90min. The isolated cells from gingiva was incubated in 37 degrees 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. GMSCs were characterized by osteogenic, adipogenic and chondrogenic differentiation, and flow cytometric, immunofluorescence analysis. ResultsGMSCs showed spindle-shaped, fibroblast-like morphology, colony-forming abilities, adherence to plastic and multilineage differentiation (osteogenic, adipogenic, chondrogenic) potency. GMSCs expressed CD44, CD73, CD90 and CD105, but did not express CD14, CD45, CD34 and CD19 in flow cytometry. Expression of stem cell markers (SSEA-4, STRO-1, CD146, CD166 and CD271) and a mesenchymal marker (vimentin) were observed by immunofluorescence. ConclusionsIn conclusion, we isolated and characterized stem cells from human gingival connective tissue with modified protocol. GMSCs showed multipotency with high proliferation and characteristics of mesenchymal stem cells. GMSCs 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 GSMCs' proliferation and cryopreservation.
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收藏
页码：461 / 467
页数：7
相关论文
共 31 条
[1]   Collagenase and dispase enzymes disrupt lymphocyte surface molecules [J].
Abuzakouk, M ;
Feighery, C ;
OFarrelly, C .
JOURNAL OF IMMUNOLOGICAL METHODS, 1996, 194 (02) :211-216
[2]   Adult human bone marrow-derived mesenchymal progenitor cells are capable of adhesion-independent survival and expansion [J].
Baksh, D ;
Davies, JE ;
Zandstra, PW .
EXPERIMENTAL HEMATOLOGY, 2003, 31 (08) :723-732
[3]   In vitro stem cell cultures from human dental pulp and periodontal ligament:: New prospects in dentistry [J].
Ballini, A. ;
De Frenza, G. ;
Cantore, S. ;
Papa, F. ;
Grano, M. ;
Mastrangelo, F. ;
Tete, S. ;
Grassi, F. R. .
INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY, 2007, 20 (01) :9-16
[4]  
Bobis S, 2006, FOLIA HISTOCHEM CYTO, V44, P215
[5]   Concise review: Mesenchymal stem cells: Their phenotype, differentiation capacity, immunological features, and potential for homing [J].
Chamberlain, Giselle ;
Fox, James ;
Ashton, Brian ;
Middleton, Jim .
STEM CELLS, 2007, 25 (11) :2739-2749
[6]   Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow [J].
Colter, DC ;
Class, R ;
DiGirolamo, CM ;
Prockop, DJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (07) :3213-3218
[7]   Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement [J].
Dominici, M. ;
Le Blanc, K. ;
Mueller, I. ;
Slaper-Cortenbach, I. ;
Marini, F. C. ;
Krause, D. S. ;
Deans, R. J. ;
Keating, A. ;
Prockop, D. J. ;
Horwitz, E. M. .
CYTOTHERAPY, 2006, 8 (04) :315-317
[8]   SSEA-4 identifies mesenchymal stem cells from bone marrow [J].
Gang, Eun J. ;
Bosnakovski, Darko ;
Figueiredo, Camila A. ;
Visser, Jan W. ;
Perlingeiro, Rita C. R. .
BLOOD, 2007, 109 (04) :1743-1751
[9]   Clinical utility of stem cells for periodontal regeneration [J].
Hynes, Kim ;
Menicanin, Danijela ;
Gronthos, Stan ;
Bartold, P. Mark .
PERIODONTOLOGY 2000, 2012, 59 :203-227
[10]   Current and future periodontal tissue engineering [J].
Izumi, Yuichi ;
Aoki, Akira ;
Yamada, Yoichi ;
Kobayashi, Hiroaki ;
Iwata, Takanori ;
Akizuki, Tatsuya ;
Suda, Tomonari ;
Nakamura, Sayaka ;
Wara-Aswapati, Nawarat ;
Ueda, Minoru ;
Ishikawa, Isao .
PERIODONTOLOGY 2000, 2011, 56 :166-187
1234