Abstract
Stem cells obtained from the body tissue, such as adipose tissue, dental pulp and gingival tissue. Fresh tissue is often used to isolate and culture for regenerative medicine. However, availability of tissue as and when required is one of the measure issue in regenerative medicine. Cryopreservation of tissue provides benefit over tissue availability, storage for significant amount of period and helps preserve the original cell structures. The effects of cryopreservation of gingival tissue for mesenchymal stem cell (MSC) are not well documented; however this process is of increasing importance for regenerative therapies. This study examined the effect of cryopreservation on the long term survival the whole gingival biopsy tissue. We studied cell outgrowth, cell morphology, MSC surface-markers and differentiation of mesenchymal stem cells derived from cryopreserved gingiva. In this study, gingival tissue was cryopreserved for 3, 6, 9 months. Cryopreserved tissue has been thawed and cells were isolated by using explant culture method. The fresh and cryopreserved gingival tissue cells were cultured and characterized for surface marker analysis, CFU-f, population doubling time, and osteogenic, chondrogenic and adipogenic differentiation. The fresh and cryopreserved tissue has similar stem cell properties. Results indicate that cryopreservation of the entire gingival tissue does not affect the properties of stem cells. This opens door for gingival tissue banking for future use in periodontology and regenerative medicine.
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References
Arutyunyan I, Fatkhudinov T, Sukhikh G (2018) Umbilical cord tissue cryopreservation: a short review. Stem Cell Res Ther 9:1–7. https://doi.org/10.1186/s13287-018-0992-0
Chen YK, Huang AHC, Chan AWS et al (2011) Human dental pulp stem cells derived from different cryopreservation methods of human dental pulp tissues of diseased teeth. J Oral Pathol Med 40:793–800. https://doi.org/10.1111/j.1600-0714.2011.01040.x
Gao Y, Zhao G, Li D et al (2014) Isolation and multiple differentiation potential assessment of human gingival mesenchymal stem cells. Int J Mol Sci 15:20982–20996. https://doi.org/10.3390/ijms151120982
Ginani F, Soares DM, Rabêlo LM et al (2016) Effect of a cryopreservation protocol on the proliferation of stem cells from human exfoliated deciduous teeth. Acta Odontol Scand 74:598–604. https://doi.org/10.1080/00016357.2016.1224919
Han YJ, Kang YH, Shivakumar SB et al (2017) Stem cells from cryopreserved human dental pulp tissues sequentially differentiate into definitive endoderm and hepatocyte-like cells in vitro. Int J Med Sci 14:1418–1429. https://doi.org/10.7150/ijms.22152
Jin SH, Lee JE, Yun JH et al (2015) Isolation and characterization of human mesenchymal stem cells from gingival connective tissue. J Periodontal Res 50:461–467. https://doi.org/10.1111/jre.12228
Kharat A, Sanap A, Kheur S et al (2022) Insulin-producing cell clusters derived from human gingival mesenchymal stem cells as a model for diabetes research. Mol Biol Rep 49:11973–11982. https://doi.org/10.1007/s11033-022-08008-6
Kim D, Lee AE, Xu Q et al (2021) Gingiva-derived mesenchymal stem cells: potential application in tissue engineering and regenerative medicine–a comprehensive review. Front Immunol 12:1–25. https://doi.org/10.3389/fimmu.2021.667221
Li J, Xu SQ, Zhang K et al (2018) Treatment of gingival defects with gingival mesenchymal stem cells derived from human fetal gingival tissue in a rat model. Stem Cell Res Ther 9:1–8. https://doi.org/10.1186/s13287-017-0751-7
Lin CS, Xin ZC, Dai J, Lue TF (2013) Commonly used mesenchymal stem cell markers and tracking labels: limitations and challenges. Histol Histopathol 28:1109–1116. https://doi.org/10.14670/HH-28.1109
Ma L, Makino Y, Yamaza H et al (2012) Cryopreserved dental pulp tissues of exfoliated deciduous teeth is a feasible stem cell resource for regenerative medicine. PLoS ONE. https://doi.org/10.1371/journal.pone.0051777
Marquez-Curtis LA, Janowska-Wieczorek A, McGann LE, Elliott JAW (2015) Mesenchymal stromal cells derived from various tissues: biological, clinical and cryopreservation aspects. Cryobiology 71:181–197
Patil VR, Kharat AH, Kulkarni DG et al (2018) Long term explant culture for harvesting homogeneous population of human dental pulp stem cells. Cell Biol Int 42:1602–1610. https://doi.org/10.1002/cbin.11065
Romanov YA, Balashova EE, Volgina NE et al (2016) Isolation of multipotent mesenchymal stromal cells from cryopreserved human umbilical cord tissue. Bull Exp Biol Med 160:530–534. https://doi.org/10.1007/s10517-016-3213-9
Seo BM, Miura M, Sonoyama W et al (2005) Recovery of stem cells from cryopreserved periodontal ligament. J Dent Res 84:907–912. https://doi.org/10.1177/154405910508401007
Sharpe PT (2016) Dental mesenchymal stem. Cells Dev 143:2273–2280. https://doi.org/10.1242/dev.134189
Tomar GB, Srivastava RK, Gupta N et al (2010) Human gingiva-derived mesenchymal stem cells are superior to bone marrow-derived mesenchymal stem cells for cell therapy in regenerative medicine. Biochem Biophys Res Commun 393:377–383. https://doi.org/10.1016/j.bbrc.2010.01.126
Wang F, Yu M, Yan X et al (2011) Gingiva-derived mesenchymal stem cell-mediated therapeutic approach for bone tissue regeneration. Stem Cells Dev 20:2093–2102. https://doi.org/10.1089/scd.2010.0523
Wang W, Yan M, Aarabi G et al (2022) Cultivation of cryopreserved human dental pulp stem cells–a new approach to maintaining dental pulp tissue. Int J Mol Sci. https://doi.org/10.3390/ijms231911485
Yan M, Nada OA, Kluwe L et al (2020) Expansion of human dental pulp cells in vitro under different cryopreservation conditions. In Vivo (brooklyn) 34:2363–2370. https://doi.org/10.21873/invivo.12049
Zhang Q, Shi S, Liu Y et al (2010) Mesenchymal stem cells derived from human gingiva are capable of immunomodulatory functions and ameliorate inflammation-related tissue destruction in experimental colitis. J Immunol 184:1656–1656. https://doi.org/10.4049/jimmunol.0990118
Zhang X, Zeng D, Huang F, Wang J (2019) A protocol for isolation and culture of mesenchymal stem cells from human gingival tissue. Am J Clin Exp Immunol 8:21–26
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Rakh, D., Kuloli, A., Kharat, A. et al. Long-term cryopreservation of whole gingival tissue. Cell Tissue Bank (2023). https://doi.org/10.1007/s10561-023-10115-y
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DOI: https://doi.org/10.1007/s10561-023-10115-y