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Abstract：
Dental follicle tissue is a promising resource of mesenchymal stem cells for cytotherapeutic approaches and tissue engineering applications. There are two procedures for banking of human dental follicle stem cells have been reported. Conventional method requires cell isolation, expansion and immediate cryopreservation. Whereas dental follicle stem cells can be isolated from cryopreserved dental follicle fragments. The aim of this study was to compare the characteristics of dental follicle cells isolated from cryopreserved fragments (DFCs-CF) with dental follicle cells recovered from cryopreserved cells (DFCs-CC). Dental follicle fragments obtained after mechanical disaggregation were divided into two parts, with one part maintained in culture, while another part underwent cryopreservation. Dental follicle fragments and dental follicle cells from fresh tissue were stored in liquid nitrogen for 3 months. After thawing, the isolation, morphology, proliferation, cell cycle, colony-forming-unit ability, stemness-related marker expression, apoptosis, and multi-lineage differentiation potential of DFCs-CF were tested compared with DFCs-CC. DFCs-CF expressed mesenchymal stem cells marker, proliferated well, showed similar levels of mRNA for stemness- and apoptosis-related genes and exhibited the capacity of multi-lineage differentiation similar to those of DFCs-CC. These results imply that cryopreservation of dental follicle fragments is an effective banking method for isolation of dental follicle cells.
摘要：
牙卵泡组织是用于细胞治疗方法和组织工程应用的间充质干细胞的有希望的资源。已经报道了两种用于保存人牙囊干细胞的程序。常规方法需要细胞隔离，扩张和立即冷冻保存。而牙囊干细胞可以从冷冻保存的牙囊碎片中分离。这项研究的目的是比较从冷冻保存的碎片（DFCs-CF）中分离的牙囊细胞与从冷冻保存的细胞（DFCs-CC）中回收的牙囊细胞的特征。将机械解聚后获得的牙卵泡碎片分为两部分，一部分保持在文化中，而另一部分接受了冷冻保存。将来自新鲜组织的牙囊碎片和牙囊细胞在液氮中储存3个月。解冻后,隔离,形态学,扩散,细胞周期,菌落形成单位能力，干性相关标记表达，凋亡，与DFCs-CC相比，检测了DFCs-CF的多谱系分化潜力。DFCs-CF表达间充质干细胞标志物，扩散良好，显示出与干细胞和凋亡相关基因的mRNA水平相似，并显示出与DFCs-CC相似的多谱系分化能力。这些结果表明，冷冻保存牙囊碎片是分离牙囊细胞的有效方法。
