口腔医学 ›› 2022, Vol. 42 ›› Issue (11): 974-978.doi: 10.13591/j.cnki.kqyx.2022.11.003

• 基础研究 • 上一篇    下一篇

基于生物打印技术的间接共培养体系的构建

毕效铭1, 王雯1,2, 高茜1, 袁长永1,3   

  1. 1 徐州医科大学口腔医学院,江苏徐州(221000);
    2 徐州市口腔医院牙周黏膜病科,江苏徐州(221000);
    3 徐州市口腔医院口腔种植科,江苏徐州(221000)
  • 收稿日期:2022-07-25 出版日期:2022-11-28 发布日期:2022-11-25
  • 通讯作者: 袁长永 Tel:(0)18952175008
  • 基金资助:
    国家自然科学基金(81700954);2020年江苏省高等学校大学生创新创业训练计划项目(202010313023Z);江苏省卫生健康委2020年度医学科研项目(M2020091)

Construction of an indirect co-culture system based on bioprinting technology

BI Xiaoming, WANG Wen, GAO Qian, YUAN Changyong   

  1. School of Stomatology, Xuzhou Medical University, Xuzhou 221000, China
  • Received:2022-07-25 Online:2022-11-28 Published:2022-11-25

摘要: 目的 构建牙髓干细胞(dental pulp stem cells, DPSCs)与人脐静脉血管内皮细胞(human umbilical vein endothelial cells, HUVECs)的间接共培养模型,为建立可应用于体内外的间接共培养体系打下基础。方法 配制5%甲基丙烯酰化明胶(GelMA)30胶,摸索三维生物打印的最佳参数,进行可打印性测试。使用活死染色实验检测DPSCs和HUVECs在GelMA30胶中的存活情况。构建6组共培养模型:DPSCs与HUVECs直接共培养(M),单纯DPSCs培养(D-D200),单纯HUVECs培养(H-D200),DPSCs与HUVECs间接共培养(M-D0、M-D200、M-D400),拍照观察细胞共存情况。结果 Pr计算结果为0.97±0.02,表明结构具有良好的可打印性。第7天DPSCs活细胞率为96.48%,HUVECs活细胞率为96.00%,证明细胞保有活力。6组共培养模型打印成功。结论 利用三维生物打印可以构建DPSCs与HUVECs的间接共培养体系,为后续的研究奠定基础。

关键词: 牙髓干细胞, 人脐静脉血管内皮细胞, 三维生物打印, 间接共培养

Abstract: Objective To constructan indirect co-culture model of dental pulp stem cells(DPSCs) and human umbilical vein endothelial cells(HUVECs), and lay a foundation for the establishment of a new indirect co-culture system that can be used in vivo and in vitro. Methods 5% GelMA30 was configured to explore the optimal parameter of 3D bioprinting and to conduct printable tests. The survival of DPSCs and HUVECs in GelMA30 was detected with live/dead staining. Six co-culture models were constructed: direct co-culture of DPSCs with HUVECs (M), culture of DPSCs(D-D200), culture of HUVECs(H-D200), indirect co-culture of DPSCs with HUVECs(M-D0, M-D200, M-D400). The co-culture of DPSCs with HUVECs was observed by photographing. Results The Pr calculation result was 0.97±0.02, indicating that the structure has good printability. On the seventh day, the viable cell rate of DPSCs and HUVECs was 96.48% and 96.00% respectively, indicating that the cells remained active. The printing of co-culture models in 6 groups was successfully completed. Conclusion Indirect co-culture system of DPSCs and HUVECs can be constructed by 3D bioprinting, which will pave the way for subsequent studies.

Key words: dental pulp stem cells, human umbilical vein endothelial cells, 3D bioprinting, indirect co-culture system

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