Simplified conditions for storing and cryopreservation of dental pulp stem cells |
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| Institution: | 1. Department of Dental Basic Sciences, Faculty of Odonto-Stomatology, University of Medicine and Pharmacy, Ho Chi Minh City, Viet Nam;2. Department of Physiology and Animal Biotechnology, Laboratory of Tissue Engineering and Biomedical Materials, Faculty of Biology – Biotechnology, University of Science, Vietnam National University, Ho Chi Minh City, Viet Nam;1. Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany;2. Institute for Biometry, Hannover Medical School, Hannover, Germany;3. Division of Hematology/Oncology, Children''s Hospital Boston, Harvard Medical School, Boston, MA, USA;1. Assistant Professor, Department of Periodontology, Faculty of Dentistry, Kirikkale University, Kirikkale, Turkey;2. Doctor, Atasehir Oral and Dental Health Clinic, Istanbul, Turkey;3. Professor, Department of Biochemistry, Faculty of Medicine, Kirikkale University, Kirikkale, Turkey;4. Doctor, Department of Biochemistry, Yildirim Beyazit University, Ankara, Turkey;6. Professor, Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Kirikkale University, Kirikkale, Turkey;1. Advanced Specialty Education Programs in Endodontics, School of Dentistry, Loma Linda University, Loma Linda, California;2. Department of Endodontics, UCLA School of Dentistry, Los Angeles, California;3. Department of Advanced Oral Sciences and Therapeutics, Division of Endodontics, School of Dentistry, University of Maryland, Baltimore, Baltimore, Maryland;4. Advanced Specialty Education Programs in Oral and Maxillofacial Surgery, School of Dentistry, Loma Linda University, Loma Linda, California;1. School of Medical Science, Australia;2. School of Medicine, Australia;3. School of Biological Sciences, University of Adelaide, Australia;4. Human Physiology, Centre for Neuroscience, School of Medicine, Flinders University, Australia;5. South Australian Health & Medical Research Institute, South Australia, Australia;1. Department of Hematology, University Medical Center Groningen, Groningen, The Netherlands;;2. Department of Hematology, HOVON Data Center, Erasmus MC Cancer Institute, Rotterdam, The Netherlands;;3. Department of Oncology, University Hospital, Inselspital, and University of Bern, Bern, Switzerland;;4. Department of Hematology, Meander Hospital Amersfoort, Amersfoort, The Netherlands;;5. Department of Hematology, Ospedale Regionale, Bellinzona, Switzerland;;6. Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius University, Vilnius, Lithuania;;7. Department of Hematology, Erasmus University Medical Center, Rotterdam, The Netherlands;;8. Department of Hematology, Amsterdam UMC, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands;;9. Department of Hematology, ZNA Stuivenberg/Middelheim, Antwerp, Belgium;;10. Department of Hematology, Hagaziekenhuis, Den Haag, The Netherlands;;11. Department of Hematology, Amphia Hospital, Breda, The Netherlands;;12. Department of Hematology, Kantonsspital St. Gallen, St. Gallen, Switzerland;;13. Department of Hematology, Medical Center Leeuwarden, Leeuwarden, The Netherlands;;14. Department of Medical Oncology and Hematology, Universitätsspital Zurich, Zurich, Switzerland;;15. Department of Hematology, Antonius Hospital, Nieuwegein, The Netherlands;;16. Department of Hematology, Isala Hospital, Zwolle, The Netherlands;;17. Department of Hematology, AZ Delta Roeselare, Roeselare, Belgium;;18. Department of Hematology, Maastricht University Medical Center, Maastricht, The Netherlands;;19. Department of Hematology, Medical Spectrum Twente, Enschede, The Netherlands;;20. Department of Hematology, Maxima Medical Center, Veldhoven, The Netherlands;;21. Division of Hematology, University Hospital Genève and Faculty of Medicine, University of Genève, Genève, Switzerland; and;22. Service and Central Laboratory of Hematology, Department of Oncology and Department of Laboratory Medicine and Pathology, Lausanne University Hospital (CHUV), Lausanne, Switzerland |
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| Abstract: | ObjectivesThis study aimed to simplify the collection, isolation and cryopreservation procedure of human dental pulp stem cells (DPSCs) to ease the establishment of dental stem cell banking.DesignExtracted third molars were collected and stored either in growth medium or in gentamicin-saline (480 μg/ml) for 6, 9 or 12 h. DPSCs were isolated and subjected to cryopreservation by a controlled-rate or rapid freezing method in 5 or 10% DMSO. Flow cytometry and growth pattern of DPSCs before and after cryopreservation were conducted.ResultsRate of contamination by which the extracted teeth were stored in control and gentamicin-saline were 9.1% (N = 33) and 2.3% (N = 43), respectively. Successful cell isolation rate of teeth preserved in gentamicin-saline at 6 h (92.9%) was comparable to those of growth media group (90.3%). At 9 and 12 h, the rates dropped significantly to 75% and 54%, respectively. Cryopreservation by controlled-rate freezing either in 5 or 10% DMSO resulted in a significantly higher percentage of viable cells than by rapid freezing. Cells conserved by controlled-rate freezing in 5% DMSO showed a pattern of growth similar to control unfrozen cells; 10% DMSO significantly deteriorated the growth pattern of the cells. After thawing, DPSCs conserved by controlled-rate freezing still expressed stemness characteristics, although hematopoietic stem cell markers were slightly increased.ConclusionGentamicin-saline was effective in preserving human teeth for DPSC isolation. Controlled-rate freezing in 5% DMSO gave the highest rate of cell viability. This study simplifies the storage conditions and proposes a simple method for cryopreservation of DPSCs. |
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| Keywords: | Dental pulp stem cells Gentamicin Storage solution Cryopreservation Cell viability |
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