| 谭金娣,何佳英,姜建平.脐带血清促进牙髓干细胞增殖作用的研究[J].浙江中西医结合杂志,2022,32(10): |
| 脐带血清促进牙髓干细胞增殖作用的研究 |
| Study 0n the Effect of Umbilical Cord Serum On the Proliferation of Dental Pulp Stem CellsTan Jindi1 ,Jiang Jianping2,He JiaYing1(Department of Hangzhou hospital of traditional ChineseMedicine ,Hangzhou310000,China; ) |
| 投稿时间：2022-04-12 修订日期：2022-09-10 |
| DOI： |
| 中文关键词: 牙髓干细胞 脐带血清 细胞培养 |
| 英文关键词:Dental pulp stem cells umbilical cord blood serum cell culture |
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| 中文摘要: |
| 目的 建立脐带血清（ umbilical cord blood serum，CBS）培养体系，替代胎牛血清（fetal bovine serum,FBS）体外扩增牙髓干细胞（dental plup stem cells,DPSC）。方法 取各项病原微生物测定皆显示阴性的人CBS用作细胞培养。提取人完整且无龋坏的正畸拔除牙或第三磨牙的牙髓干细胞，设置四个实验组，其中对照组为含10%胎牛血清培养基培养的牙髓干细胞，其他三组分别为含5%、10%、20%脐带血清培养基培养的牙髓干细胞。经由MTT法对比4个组细胞增殖速度；对增殖速度最快的一组，通过不同诱导液对其DPSCs施以诱导，使其分别分化为脂肪细胞、成骨细胞、成软骨细胞，对其多向分化活性加以证实；FCM（流式细胞术）对细胞表面标记物表达展开测定。结果1、脐带血清可培养出能够稳定传代的牙髓干细胞系；2、MTT结果显示，各组牙髓干细胞在1，3，5，7，9天内均呈现不同程度的生长状态，且呈线性增长，呈“S”型。其中10%脐带血清组在第7天后细胞增殖较为明显；3、培养出的牙髓干细胞具有多向分化能力；4、FCM测定培养出的第3代DPSC表面标记物表达水平，MSCA-1、CD73与CD90为阳性表达，CD34与CD45为阴性表达，与间充质干细胞表面标记物表达特性相符。结论 脐带血清培养体系于体外条件下对DPSC增殖具促进作用，且培养所得DPSC具备干细胞特性。 |
| 英文摘要: |
| Abstract Objective The establishment of a cord blood serum (CBS) culture system to replace fetal bovine serum (FBS) for in vitro expansion of dental pulp stem cells (DPSC). Methods For cell culture, healthy human CBS that are negative for all pathogenic microorganisms are taken. After extracting DPSC from orthodontically extracted teeth or third molars in healthy adults without caries, four experimental groups are established, in which the control group is DPSC cultured with 10% FBS medium, and the other three groups are DPSC cultured with 5%, 10% and 20% CBS medium, respectively. After comparing the proliferation rate of the four groups by MTT, the fastest proliferating group is induced with different induction solutions to differentiate into adipocytes, osteoblasts and chondrocytes to confirm their multidirectional differentiation activity. Cell surface marker expression is determined by flow cytometry (FCM). Results Umbilical cord serum can be grown to produce DPSC capable of stable transmission. The MTT results show that all groups of dental pulp stem cells present different degrees of growth at 1, 3, 5, 7 and 9 d. The growth is linear and "S" shaped. In which the cell proliferation in the 10% CBS group is more obvious after the 7th d. Cultivated DPSC has multidirectional differentiation ability. When FCM measured the expression levels of surface markers of cultured 3rd generation DPSC, MSCA-1, CD73 and CD90 are positive and CD34 and CD45 are negative, which are consistent with the surface marker expression characteristics of MSCs. Conclusion Under the in vitro conditions, the CBS culture system can promote the proliferation of DPSC and the resulting DPSC have stem cell properties.
Keywords dental pulp stem cells; umbilical cord serum; cell culture; cell proliferation |
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