title: Cryopreservation of human dental follicle tissue for use as a resource of autologous mesenchymal stem cells.
abstract: The main purpose of this study was to develop a cryopreservation method for human dental follicle tissue to maintain autologous stem cells as a resource. A modified cryoprotectant, consisting of 0.05 m glucose, 0.05 m sucrose and 1.5 m ethylene glycol in phosphate-buffered saline (PBS) was employed, with a slow-ramp freezing rate. We observed > 70% of cell survival rate after 3 months of tissue storage. Isolated and cultured human dental stem cells (hDSCs) from cryopreserved dental follicles expressed mesenchymal stem cell markers at a level similar to that of hDSCs from fresh tissue. They also successfully differentiated in vitro into the mesenchymal lineage, osteocytes, adipocytes and chondrocytes under specific inductions. Using immunohistochemistry, the early transcription factors OCT4, NANOG and SOX2 were moderately or weakly detected in the nucleus of both fresh and cryopreserved dental follicles. In addition, p63, CCND1, BCL2 and BAX protein expression levels were the same in both fresh and cryopreserved tissues. However, the positive-cell ratio and intensity of p53 protein was higher in cryopreserved tissues than in fresh tissues, indicating direct damage of the freeze-thawing process. Real-time PCR analysis of hDSCs at passage 2 from both fresh and cryopreserved dental follicles showed similar levels of mRNA for apoptosis- and transcription-related genes. Based on these results, a newly developed cryoprotectant, along with a slow ramp rate freezing procedure allows for long-term dental tissue preservation for later use as an autologous stem cell resource in regenerative cell therapy. Copyright © 2014 John Wiley & Sons, Ltd.
authors: Department of Oral and Maxillofacial Surgery, School of Medicine and Institute of Health Science, Gyeongsang National University, Jinju, Republic of Korea.
authors: Bong-Wook
authors: BW
authors: Park
authors: OBS/Theriogenology and Biotechnology, Gyeongsang National University, Jinju, Republic of Korea.
authors: Si-Jung
authors: SJ
authors: Jang
authors: Department of Oral and Maxillofacial Surgery, School of Medicine and Institute of Health Science, Gyeongsang National University, Jinju, Republic of Korea.
authors: June-Ho
authors: JH
authors: Byun
authors: Department of Oral and Maxillofacial Surgery, School of Medicine and Institute of Health Science, Gyeongsang National University, Jinju, Republic of Korea.
authors: Young-Hoon
authors: YH
authors: Kang
authors: Department of Oral and Maxillofacial Surgery, School of Medicine and Institute of Health Science, Gyeongsang National University, Jinju, Republic of Korea.
authors: Mun-Jeong
authors: MJ
authors: Choi
authors: Department of Ceramic Engineering, Gyeongsang National University, Jinju, Republic of Korea.
authors: Won-Uk
authors: WU
authors: Park
authors: OBS/Theriogenology and Biotechnology, Gyeongsang National University, Jinju, Republic of Korea.
authors: Won-Jae
authors: WJ
authors: Lee
authors: OBS/Theriogenology and Biotechnology, Gyeongsang National University, Jinju, Republic of Korea.|Research Institute of Life Sciences, Gyeongsang National University, Jinju, Republic of Korea.
authors: Gyu-Jin
authors: GJ
authors: Rho
mesh_terms: D017667:Adipocytes / Q000166:cytology; D000293:Adolescent; D000328:Adult; D017209:Apoptosis; D019070:Cell Lineage; D002470:Cell Survival; D019902:Chondrocytes / Q000166:cytology; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000737:chemistry; D003795:Dental Sac / Q000166:cytology* / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008964:Molar, Third / Q000166:cytology; D010011:Osteocytes / Q000166:cytology; D023822:Tissue Engineering; D055815:Young Adult
chemical_list: D003451:Cryoprotective Agents
keywords: autologous stem cell source; human dental follicle tissue; mesenchymal stem cells (MSCs); regenerative medicine; tissue cryopreservation