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The establishment of a chemically defined serum-free culture system for human dental pulp stem cells

机译:建立化学定义的人牙髓干细胞无血清培养系统

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摘要

Abstract Background The concept of establishing a dental stem cell (DSC) bank for oral and maxillofacial regeneration has become of great interest but it remains at a primitive stage. The routine application of serum-containing conditions for human DSC (hDSC) culture is in great controversy considering that the animal-originated serum can cause serious ethical concerns and lead to increasingly irrelevant variables, errors, and poor repeatability of experiment results. Thus, this study aimed to establish a safe, stable and efficient hDSC serum-free culturing system for future DSC bank usage. Methods Dental pulp stem cells (DPSCs) from human permanent tooth pulp were isolated, expanded, passaged, and divided into two groups according to their culture conditions: group 1 was the serum-containing medium (SCM) group; and group 2 was the serum-free Essential 8 medium (E8) group. DPSCs were characterized first, followed by cell proliferation, pluripotency, and migration study in SCM and E8 medium. Results Human DPSCs (hDPSCs) in E8 medium demonstrated greater proliferation, pluripotency, migration ability and less apoptosis. hDPSCs could be successfully induced to the adipogenic, osteogenic, neurogenic, and chondrogenic lineages in E8 group. Real-time polymerase chain reaction indicated that the expression of PPAR-γ, RUNX2, OCN and MAP-2 was higher in E8 group.  Conclusions Compared with serum-containing medium, E8 medium exhitibed higher ability in maintaining the cell proliferation, pluripotency, migration, and stability. This new serum-free culture environment might be applicable for hDSC culture in the future.
机译:摘要背景建立牙齿干细胞(DSC)银行为口腔和颌面复再生的概念变得非常兴趣,但它仍处于原始阶段。考虑到动物起源的血清可能导致严重的道德问题并导致越来越多的变量,误差和实验结果的可重复性越来越多的伦理问题,含血清患者的血清培养物(HDSC)培养物的常规应用具有很大的争议。因此,本研究旨在为未来的DSC银行使用建立一种安全,稳定和高效的HDSC血清培养系统。方法根据其培养条件分离,膨胀,传代,分离来自人永齿纸浆的牙科纸浆干细胞(DPSC),并分为两组:第1组是含血清培养基(SCM)组;第2组是无血清必需品8中(E8)组。首先表征了DPSC,其次是SCM和E8培养基中的细胞增殖,多能性和迁移研究。结果E8培养基中的人DPSCs(HDPSC)显示出更大的增殖,多能性,迁移能力和更少的细胞凋亡。 HDPSC可以成功地诱导E8组中的脂肪发生,成骨,神经发生和软骨内谱系。实时聚合酶链反应表明,在E8组中,PPAR-γ,runx2,OCN和MAP-2的表达更高。结论与含血清培养基相比,E8培养基对维持细胞增殖,多能性,迁移和稳定性进行了更高的能力。这种无血清文化环境可能适用于未来的HDSC文化。

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