| 王 甜,郭 玲,安广洲,周 艳,张俊平,张克英,郭国祯,丁桂荣.1950 MHz射频电磁场对人脐带间充质干细胞增殖和成骨分化的影响研究[J].,2018,(12):2218-2223 |
| 1950 MHz射频电磁场对人脐带间充质干细胞增殖和成骨分化的影响研究 |
| Effects of 1950 MHz Radiofrequency Electromagnetic Fields on the Proliferation and Osteogenic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells |
| 投稿时间：2017-12-08 修订日期：2018-01-12 |
| DOI：10.13241/j.cnki.pmb.2018.12.004 |
| 中文关键词: 射频电磁场 间充质干细胞 细胞增殖 成骨分化 |
| 英文关键词: Radiofrequency electromagnetic fields Mesenchymal stem cells Cell proliferation Osteogenic differentiation |
| 基金项目:国家自然科学基金项目（31770905） |
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| 中文摘要: |
| 摘要 目的：间充质干细胞(Mesenchymal stem cells，MSCs）具有广阔的临床应用前景，但由于其体外增殖和定向分化等问题，制约了其进一步应用。本研究拟探讨1950MHz射频电磁场（Radio-frequency electromagnetic fields, RF-EMF）对人脐带间充质干细胞（Human umbilical cord mesenchymal stem cells, hUC-MSCs）增殖和成骨方向分化的影响，以期为MSCs的体外增殖和定向分化提供一条新途径。方法：华通氏胶组织块法分离培养人脐带间充质干细胞，流式细胞仪检测间充质干细胞特异性标志物。选择鉴定后的第3至第6代（P3-P6）hUC-MSCs用于实验。将hUC-MSCs细胞暴露或假暴露于频率为1950 MHz，比吸收率（Specific ab- sorption rate, SAR）分别为0.5，1.0和2.0 W/kg的RF-EMF中，每天暴露1 h（5 min开，10 min关），连续暴露7 d。暴露结束后，流式细胞仪检测细胞周期，免疫荧光检测增殖相关蛋白Ki67表达，连续6天用CCK-8方法检测细胞数。在成骨分化研究中，将P3代的hUC-MSCs随机分为假暴露（sham）组，射频辐射暴露（RF）组，成骨诱导培养基组（Induction medium, OM）和成骨诱导培养基联合射频辐射暴露（OM+RF）组，暴露SAR值为2.0 W/kg，其它参数不变。暴露结束后立即检测细胞的碱性磷酸酶（Alkaline phos- phatase, ALP）活性。结果：原代培养的细胞具有MSC典型外观，且表达MSCs特异性表面抗原。与sham组相比，不同SAR值RF暴露后，hUC-MSCs的增殖能力无明显变化，S期细胞比例及Ki67蛋白水平也无显著改变。此外，hUC-MSCs经SAR值为2.0 W/kg的RF暴露7 d，与sham组相比其ALP活性无显著变化。与OM组相比，OM+RF组的ALP活性亦无显著改变。结论：华通氏胶组织块法能够培养出纯度较高的间充质干细胞，本实验条件下的1950 MHz射频电磁场对hUC-MSCs的增殖和成骨分化均无显著影响。 |
| 英文摘要: |
| ABSTRACT Objective: Mesenchymal stem cells (MSCs) has wide prospect of clinical application, but its proliferation and direc- tional differentiation in vitro restrict its further application. In the study, we explored the effects of 1950 MHz radiofrequency electromag- netic fields (RF-EMF) on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs), in order to provide a new way for the proliferation and differentiation of MSCs in vitro. Methods: The hUC-MSCs were cultured by ex- plant method of Wharton's jelly and the specific markers of mesenchymal stem cells were detected by flow cytometry(FCM). The hUC-MSCs of passage 3 to 6 were assigned and intermittent exposed or sham-exposed to 1950 MHz RF-EMF at specific absorption rates (SARs) of 0.5, 1.0 or 2.0 W/kg for 7 consecutive days (1 h/d), 5 min on and 10 min off. After exposure, cell viability was detected by CCK-8, cell cycle was measured by flow cytometry, and Ki67 protein expression was tested by immunofluorescence. In the study of os- teogenic differentiation, the hUC-MSCs of passage 3 were divided into sham group, RF group, induction medium (OM) group and OM+RF group. After the same exposure above, except the SAR is 2.0 W/kg, the alkaline phosphatase (ALP) activity was measured im- mediately. Results: The cells cultured by explant method showed typical appearance of MSCs, and expressed MSCs specific surface anti- gen positively. Compared with sham-exposed cells, the proliferation capacity of hUC-MSCs did not change significantly after RF-EMF exposure at different SAR values, and the percentage of cells in S phase and the level of Ki67 protein expression also did not change obviously.After exposure, the activity of ALP between sham group and RF group, as well as OM and OM+RF group, had no significant dif- ferences. Conclusion: The hUC-MSCs cells could be cultured by the method of Wharton's jelly explant with high purity. 1950 MHz RF-EMF in the experiment had no significant effects on the proliferation and osteogenic differentiation of hUC-MSCs in vitro. |
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