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Effects and mechanisms of basic fibroblast growth factor on the proliferation and regenerative profiles of cryopreserved dental pulp stem cells
Cell Proliferation ( IF 8.5 ) Pub Date : 2020-12-17 , DOI: 10.1111/cpr.12969
Lihua Luo 1 , Yanni Zhang 1 , Hongyu Chen 2 , Fengting Hu 1 , Xiaoyan Wang 1 , Zhenjie Xing 1 , Abdullkhaleg Ali Albashari 1 , Jian Xiao 3 , Yan He 4 , Qingsong Ye 1, 5
Affiliation  

OBJECTIVES Various factors could interfere the biological performance of DPSCs during post-thawed process. Yet, little has been known about optimization of the recovery medium for DPSCs. Thus, our study aimed to explore the effects of adding recombinant bFGF on DPSCs after 3-month cryopreservation as well as the underlying mechanisms. MATERIALS AND METHODS DPSCs were extracted from impacted third molars and purified by MACS. The properties of CD146+ DPSCs (P3) were identified by CCK-8 and flow cytometry. After cryopreservation for 3 months, recovered DPSCs (P4) were immediately supplied with a series of bFGF and analysed cellular proliferation by CCK-8. Then, the optimal dosage of bFGF was determined to further identify apoptosis and TRPC1 channel through Western blot. The succeeding passage (P5) from bFGF pre-treated DPSCs was cultivated in bFGF-free culture medium, cellular proliferation and stemness were verified, and pluripotency was analysed by neurogenic, osteogenic and adipogenic differentiation. RESULTS It is found that adding 20 ng/mL bFGF in culture medium could significantly promote the proliferation of freshly thawed DPSCs (P4) through suppressing apoptosis, activating ERK pathway and up-regulating TRPC1. Such proliferative superiority could be inherited to the succeeding passage (P5) from bFGF pre-stimulated DPSCs, meanwhile, stemness and pluripotency have not been compromised. CONCLUSIONS This study illustrated a safe and feasible cell culture technique to rapidly amplify post-thawed DPSCs with robust regenerative potency, which brightening the future of stem cells banking and tissue engineering.

中文翻译:

碱性成纤维细胞生长因子对冷冻保存牙髓干细胞增殖和再生特性的影响及机制

目的 在解冻后的过程中,多种因素可能会干扰 DPSCs 的生物学性能。然而,关于 DPSCs 恢复介质的优化知之甚少。因此,我们的研究旨在探讨添加重组 bFGF 对冷冻保存 3 个月后 DPSCs 的影响及其潜在机制。材料与方法 从受影响的第三磨牙中提取 DPSCs 并通过 MACS 纯化。CD146+ DPSCs (P3) 的特性通过 CCK-8 和流式细胞术鉴定。冷冻保存 3 个月后,立即向回收的 DPSCs (P4) 提供一系列 bFGF,并通过 CCK-8 分析细胞增殖。然后,通过蛋白质印迹确定bFGF的最佳剂量以进一步鉴定细胞凋亡和TRPC1通道。来自 bFGF 预处理的 DPSC 的后续传代 (P5) 在不含 bFGF 的培养基中培养,验证细胞增殖和干性,并通过神经源性、成骨性和脂肪源性分化分析多能性。结果发现在培养基中加入20 ng/mL bFGF可通过抑制细胞凋亡、激活ERK通路和上调TRPC1,显着促进刚解冻的DPSCs(P4)的增殖。这种增殖优势可以遗传给 bFGF 预刺激 DPSC 的后续传代 (P5),同时,干性和多能性没有受到损害。结论 本研究展示了一种安全可行的细胞培养技术,可快速扩增具有强大再生能力的解冻后 DPSC,这为干细胞库和组织工程的未来带来光明。
更新日期:2020-12-17
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