Marędziak Monika, Marycz Krzysztof, Smieszek Agnieszka, Lewandowski Daniel, Toker Nezir Yaşar
Electron Microscopy Laboratory, University of Environmental and Life Sciences Wroclaw, Kozuchowska 5b, 51-631, Wroclaw, Poland,
In Vitro Cell Dev Biol Anim. 2014 Jun;50(6):562-71. doi: 10.1007/s11626-013-9730-1. Epub 2014 Jan 30.
The aim of this study was to evaluate the proliferation rate and morphological changes of adipose-derived mesenchymal stem cells of canine and equine origin (Eq- and CaAdMSC). Investigated cells were exposed to a static magnetic field (MF) with the intensity of 0.5 T. Proliferation activity of cells was determined with the Alamar Blue assay. Obtained results, normalized in respect to the control culture, showed that EqAdMSC exposed to MF maintained a high proliferation status, whereas proliferation activity of CaAdMSC cultured in the presence of MF was decreased. Estimations of population doubling time (PDT) also revealed that EqAdMSCs exposed to static MF achieved a twofold increase in the total number of cells in a shorter amount of time than the control culture. The PDT value obtained for investigated CaAdMSCs indicated that MF exposure resulted in the prolongation of population doubling time. Morphology of cells and cellular composition was investigated using a light inverted microscope and a fluorescent microscope. A scanning electron microscope was used for microvesicles (MVs) imaging. Obtained results showed that both cell types maintained fibroblastic morphology and did not reveal signs of apoptosis or necrosis. However, the MF had an influence on the MVs secretion. While EqAdMSCs propagated in the presence of MF were characterized by the abundant MVs presence, CaAdMSCs revealed poor secretory activity. The approach presented provides complex analysis, which enables one to determine changes in equine and canine cytophysiology.
本研究的目的是评估犬源和马源脂肪间充质干细胞(Eq-和CaAdMSC)的增殖率和形态变化。将研究的细胞暴露于强度为0.5 T的静磁场(MF)中。用阿拉玛蓝法测定细胞的增殖活性。相对于对照培养物进行标准化处理后得到的结果表明,暴露于MF的EqAdMSC保持较高的增殖状态,而在MF存在下培养的CaAdMSC的增殖活性降低。群体倍增时间(PDT)的估计还表明,暴露于静磁场的EqAdMSC在比对照培养物更短的时间内使细胞总数增加了两倍。所研究的CaAdMSC获得的PDT值表明,暴露于MF导致群体倍增时间延长。使用倒置光学显微镜和荧光显微镜研究细胞形态和细胞组成。用扫描电子显微镜对微泡(MVs)进行成像。获得的结果表明,两种细胞类型均保持成纤维细胞形态,未显示凋亡或坏死迹象。然而,MF对MVs的分泌有影响。在MF存在下增殖的EqAdMSC的特点是存在大量MVs,而CaAdMSC的分泌活性较差。所提出的方法提供了综合分析,能够确定马和犬细胞生理学的变化。