兔牙髓干细胞与雪旺细胞体外间接共培养对增殖和兔牙髓干细胞成神经向分化的研究

doi:10.13701/j.cnki.kqyxyj.2020.10.018

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (10): 968-972.DOI: 10.13701/j.cnki.kqyxyj.2020.10.018

兔牙髓干细胞与雪旺细胞体外间接共培养对增殖和兔牙髓干细胞成神经向分化的研究

张维甲, 潘爽, 李艳萍, 何丽娜, 吴作栋, 牛玉梅^*

哈尔滨医科大学附属第一医院哈尔滨医科大学口腔医学院黑龙江哈尔滨 150001

收稿日期:2020-01-22 出版日期:2020-10-28 发布日期:2020-10-19
通讯作者: *牛玉梅,E-mail:yumeiniu@163.com
作者简介:张维甲(1994~ ),男,山东青岛人,硕士,医师,主要从事牙体牙髓病科临床治疗工作。
基金资助:
国家自然科学基金(编号:81970924);中国博士后基金(编号:2018M641871);黑龙江省博士后基金(编号:LBH-Z17177)

Effect of In Vitro Indirect Co-culture of Rabbit Dental Pulp Stem Cells with Schwann Cells on Proliferation and Neuronal Differentiation of Rabbit Dental Pulp Stem Cells

ZHANG Weijia, PAN Shuang, LI Yanping, HE Lina, WU Zuodong, NIU Yumei^*

Department of Endodontics, Stomatological Hospital, Harbin Medical University, Harbin 150001, China

Received:2020-01-22 Online:2020-10-28 Published:2020-10-19

摘要/Abstract

摘要： 目的: 体外间接共培养兔牙髓干细胞(rabbit dental pulp stems cells,rDPSCs)与兔雪旺细胞(rabbit Schwann cells, rSCs),研究两种细胞对彼此增殖的影响以及rSCs诱导rDPSCs神经向分化的可能性。方法: 通过Transwell间接共培养系统构建rSCs/rDPSCs间接共培养体系,采用MTT法检测两种细胞对彼此增殖的影响;通过免疫荧光染色和Western bolt检测S-100β和NF-H的表达,研究rSCs诱导rDPSCs神经向分化的可能性。结果: 将rDPSCs与rSCs以1∶2比例间接共培养5 d时,rDPSCs能够促进rSCs的增殖(P<0.05),同时rSCs对rDPSCs的增殖也有明显的促进作用(P<0.05);在与rSCs间接共培养7 d与14 d后,rDPSCs形态与神经细胞类似,并表达S-100β和NF-H,并且14 d时两种蛋白表达更高(P<0.01)。结论: 将rDPSCs与rSCs间接共培养,5 d时rDPSCs与rSCs能显著促进彼此的增殖,且rSCs能诱导rDPSCs神经向分化。

关键词: 兔牙髓干细胞, 兔雪旺细胞, 间接共培养, 细胞增殖, 成神经向分化

Abstract: Objective: To study the effects of indirect co-culture of rabbit dental pulp stem cells (rDPSCs) and rabbit schwann cells (rSCs) in vitro on each other's proliferation and the possibility of rSCs inducing rDPSCs into neural differentiation. Methods: The indirect co-culture system of rSCs/rDPSCs was constructed by Transwell indirect co-culture system, and MTT method was adopted to detect the cells' proliferation. Immunofluorescence staining and Western bolt were used to study the expression of S-100β and NF-H and the possibility of rSCs to induce neural differentiation of rDPSCs. Results: When rDPSCs and rSCs were indirectly co-cultured in a 1∶2 ratio for 5 days, rDPSCs could facilitate the proliferation of rSCs (P<0.05), and rSCs also had obvious effect on the proliferation of rDPSCs (P<0.05). After indirect co-culturet with rSCs for 7 days and 14 days, rDPSCs became neuron-like cells, S-100β and NF-H were expressed and both presented a higher level after 14 days (P<0.01). Conclusion: In the indirect co-culture system of rDPSCs and rSCs, rDPSCs and rSCs can significantly promote each other's proliferation after 5 days, while rSCs can induce the neural differentiation of rDPSCs.

Key words: rabbit dental pulp stem cells, rabbit Schwann cells, transwell, cell proliferation, neural differentiation

张维甲, 潘爽, 李艳萍, 何丽娜, 吴作栋, 牛玉梅. 兔牙髓干细胞与雪旺细胞体外间接共培养对增殖和兔牙髓干细胞成神经向分化的研究[J]. 口腔医学研究, 2020, 36(10): 968-972.

ZHANG Weijia, PAN Shuang, LI Yanping, HE Lina, WU Zuodong, NIU Yumei. Effect of In Vitro Indirect Co-culture of Rabbit Dental Pulp Stem Cells with Schwann Cells on Proliferation and Neuronal Differentiation of Rabbit Dental Pulp Stem Cells[J]. Journal of Oral Science Research, 2020, 36(10): 968-972.

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兔牙髓干细胞与雪旺细胞体外间接共培养对增殖和兔牙髓干细胞成神经向分化的研究

Effect of In Vitro Indirect Co-culture of Rabbit Dental Pulp Stem Cells with Schwann Cells on Proliferation and Neuronal Differentiation of Rabbit Dental Pulp Stem Cells

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