优化细胞培养中牙髓细胞球体形成的条件。
Optimizing Conditions for Spheroid Formation of Dental Pulp Cells in Cell Culture.
机构信息
Department of Oral and Craniomaxillofacial Surgery, University Hospital Hamburg-Eppendorf, Hamburg, Germany.
Department of Neurology, University Hospital Hamburg-Eppendorf, Hamburg, Germany.
出版信息
In Vivo. 2021 Jul-Aug;35(4):1965-1972. doi: 10.21873/invivo.12464.
BACKGROUND/AIM: Spheroid formation is a well-known feature of stem/progenitor cells. Dental pulp cells (DPCs) cultured in serum-free medium can also form spheroids. However, the success rate varies largely depending on various factors. This study aimed to explore these factors and optimize the conditions.
MATERIALS AND METHODS
Primary DPCs were obtained from 6 wisdom teeth. Possible influencing factors including donor teeth, concentrations of the KnockOut Serum Replacement (KSR), seeding density (regarding surface and volume), passage and freezing were tested.
RESULTS
DPCs from all 6 donor teeth formed spheroids in serum-free medium. Number, size, and total area of spheroids varied among different donor teeth. Optimal concentration of the KSR and seeding densities also varied from tooth to tooth. Generally, high KSR and high cell density lead to better spheroid formation. However, very high KSR and cell density can also lead to cell death and the fusion of spheroids to irregular aggregates.
CONCLUSION
An initial setting can be recommended as: Serum-free MEM plus 10-15% KSR and seeding densities of 1-2×10 cells/ml and 2×10 cells/cm These parameters provide a direction for optimizing the condition for obtaining spheroids from human DPCs.
背景/目的：球体形成是干细胞/祖细胞的一个显著特征。无血清培养基中培养的牙髓细胞（DPCs）也可以形成球体。然而，成功率因各种因素而有很大差异。本研究旨在探讨这些因素并优化条件。
材料和方法
从 6 颗智齿中获得原代 DPCs。可能的影响因素包括供体牙齿、KnockOut Serum Replacement（KSR）的浓度、接种密度（关于表面和体积）、传代和冷冻。
结果
来自所有 6 个供体牙齿的 DPCs 在无血清培养基中形成球体。不同供体牙齿的球体数量、大小和总面积不同。KSR 的最佳浓度和接种密度也因牙齿而异。通常，高 KSR 和高细胞密度导致更好的球体形成。然而，非常高的 KSR 和细胞密度也会导致细胞死亡和球体融合成不规则的聚集体。
结论
可以推荐一个初始设置：无血清 MEM 加 10-15% KSR 和接种密度为 1-2×10 个细胞/ml 和 2×10 个细胞/cm。这些参数为优化从人牙髓细胞获得球体的条件提供了方向。
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