Cultivation and expansion of mesenchymal stem cells from human gingival tissue and periodontal ligament in different culture media

Banun Kusumawardani, Dwi Merry Christmarini Robin, Endah Puspitasari, Irma Josefina Savitri, Dea Ajeng Pravita Suendi

= http://dx.doi.org/10.20473/j.djmkg.v54.i1.p39-45
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Abstract


Background: Gingival tissue and periodontal ligament act as sources of mesenchymal stem cells (MSCs) that play a vital role in periodontal regeneration, but they both have limitations for cell availability. MSCs cultivated and expanded in various media formulations could be used as a basis for the development of cell therapy protocols. Purpose: This study aimed to determine the optimum culture media formulation for cultivation and expansion of human gingival-derived mesenchymal stem cells (hGMSCs) and human periodontal ligament stem cells (hPDLSCs). Methods: The hGMSCs and hPDLSCs were obtained from gingival tissue and periodontal ligament specimens from an adult patient. The two different culture media formulations used were: 1) α-minimum essential media (α-MEM) supplemented with 10% FBS, 100 U/mL penicillin, 100mg/mL streptomycin and 2.5 µg/mL amphotericin B; and 2) Dulbecco’s minimum essential media-Low Glucose (DMEM-LG) supplemented with 10% FBS, 2 mMol/L L-glutamine, 100 U/mL penicillin, 100mg/mL streptomycin and 2.5 µg/mL amphotericin B. The minced-gingival tissue and periodontal ligament samples were seeded in 3 cm tissue culture dishes with one of two experimental culture media, and incubated at 37oC in a humidified atmosphere of 5% CO2. Results: Cell morphology was observed on days two and five of the third passage. The gingival tissue and periodontal ligament primary cells exhibited fibroblast-like morphology, long processes and were spindle-shaped. The hPDLSCs grown in α-MEM exhibited a significant increase in cell viability and proliferation rate compared to the hPDLSCs grown in DMEM-LG. However, hGMSCs displayed similar cell viability and proliferation rate on both types of experimental media. Both the hGMSCs and hPDLSCs expressed MSC markers, including CD105, CD146, and CD90, but did not express CD45. Conclusion: Culture media formulations of α-MEM and DMEM-LG can be used for the cultivation and expansion of both hGMSCs and hPDLSCs.

Keywords


cultivation; expansion; gingival-derived mesenchymal stem cells; periodontal ligament stem cells; proliferation rate

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Yang JW, Shin YY, Seo Y, Kim H-S. Therapeutic functions of stem sells from oral cavity: an update. Int J Mol Sci. 2020; 21(12): 4389.

Zhu W, Liang M. Periodontal ligament stem cells: current status, concerns, and future prospects. Stem Cells Int. 2015; 2015: 1–11.

Assem M, Kamal S, Sabry D, Soliman N, Aly RM. Preclinical assessment of the proliferation capacity of gingival and periodontal ligament stem cells from diabetic patients. Open access Maced J Med Sci. 2018; 6(2): 254–9.

Lang NP, Bartold PM. Periodontal health. J Clin Periodontol. 2018; 45(Suppl 20): S9–16.

Cheng K-H, Kuo T-L, Kuo K-K, Hsiao C-C. Human adipose-derived stem cells: Isolation, characterization and current application in regeneration medicine. Genomic Med Biomarkers, Heal Sci. 2011; 3(2): 53–62.

McCormack SW, Witzel U, Watson PJ, Fagan MJ, Gröning F. The biomechanical function of periodontal ligament fibres in orthodontic tooth movement. PLoS One. 2014; 9(7): e102387.

Alipour F, Parham A, Kazemi Mehrjerdi H, Dehghani H. Equine adipose-derived mesenchymal stem cells: phenotype and growth characteristics, gene expression profile and differentiation potentials. Cell J. 2015; 16(4): 456–65.

Jung I-H, Kwon B-S, Kim S-H, Shim H-E, Jun C-M, Yun J-H. Optimal medium formulation for the long-term expansion and maintenance of human periodontal ligament stem cells. J Periodontol. 2013; 84(10): 1434–44.

Nirmala MJ, Durai L, Gopakumar V, Nagarajan R. Preparation of celery essential oil-based nanoemulsion by ultrasonication and evaluation of its potential anticancer and antibacterial activity. Int J Nanomedicine. 2020; 15: 7651–66.

El Fattah MA, Ding G, Wei F, Zhang C, Ezz EA, Wang S. Identification and cementoblastic / osteoblastic differentiation of postnatal stem cells from human periodontal ligament. Maced J Med Sci. 2011; 4(1): 37–43.

Bartold PM, Gronthos S. Standardization of criteria defining periodontal ligament stem cells. J Dent Res. 2017; 96(5): 487–90.

Iwata T, Yamato M, Zhang Z, Mukobata S, Washio K, Ando T, Feijen J, Okano T, Ishikawa I. Validation of human periodontal ligament-derived cells as a reliable source for cytotherapeutic use. J Clin Periodontol. 2010; 37(12): 1088–99.

Salehinejad P, Alitheen NB, Nematollahi-Mahani SN, Ali AM, Omar AR, Janzamin E, Hajghani M. Effect of culture media on expansion properties of human umbilical cord matrix-derived mesenchymal cells. Cytotherapy. 2012; 14(8): 948–53.

Yuan X, Logan TM, Ma T. Metabolism in human mesenchymal stromal cells: A missing link between hMSC biomanufacturing and therapy? Front Immunol. 2019; 10: 977.

Sánchez-Kopper A, Becker M, Pfizenmaier J, Kessler C, Karau A, Takors R. Tracking dipeptides at work-uptake and intracellular fate in CHO culture. AMB Express. 2016; 6: 48.

Petrenko Y, Vackova I, Kekulova K, Chudickova M, Koci Z, Turnovcova K, Kupcova Skalnikova H, Vodicka P, Kubinova S. A comparative analysis of multipotent mesenchymal stromal cells derived from different sources, with a focus on neuroregenerative potential. Sci Rep. 2020; 10: 4290.

Viswanathan S, Shi Y, Galipeau J, Krampera M, Leblanc K, Martin I, Nolta J, Phinney DG, Sensebe L. Mesenchymal stem versus stromal cells: International Society for Cell & Gene Therapy (ISCT®) mesenchymal stromal cell committee position statement on nomenclature. Cytotherapy. 2019; 21(10): 1019–24.

Soares IMV, Fernandes GV de O, Larissa Cordeiro C, Leite YKP de C, Bezerra D de O, Carvalho MAM de, Carvalho CMRS. The influence of Aloe vera with mesenchymal stem cells from dental pulp on bone regeneration: characterization and treatment of non-critical defects of the tibia in rats. J Appl Oral Sci. 2019; 27: e20180103.

Mrozik K, Gronthos S, Shi S, Bartold PM. A method to isolate, purify, and characterize human periodontal ligament stem cells. Methods Mol Biol. 2017; 1537: 413–27.

Hung BP, Hutton DL, Kozielski KL, Bishop CJ, Naved B, Green JJ, Caplan AI, Gimble JM, Dorafshar AH, Grayson WL. Platelet-derived growth factor BB enhances osteogenesis of adipose-derived but not bone marrow-derived mesenchymal stromal/stem cells. Stem Cells. 2015; 33(9): 2773–84.

Lei M, Li K, Li B, Gao L-N, Chen F-M, Jin Y. Mesenchymal stem cell characteristics of dental pulp and periodontal ligament stem cells after in vivo transplantation. Biomaterials. 2014; 35(24): 6332–43.

Iwasaki K, Komaki M, Yokoyama N, Tanaka Y, Taki A, Kimura Y, Takeda M, Oda S, Izumi Y, Morita I. Periodontal ligament stem cells possess the characteristics of pericytes. J Periodontol. 2013; 84(10): 1425–33.


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