Centre of Biological Engineering, Wolfson School of Mechanical & Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK.
JPK Instruments Limited, Unit 7223, Cambridge Research Park, Waterbeach, Cambridge CB25 9TL, UK.
J Biomech. 2014 Feb 7;47(3):625-30. doi: 10.1016/j.jbiomech.2013.12.004. Epub 2013 Dec 12.
This note reports observations of the change of stiffness of human mesenchymal stem cells (hMSCs) with the progress of cell death as measured by AFM. hMSC with impaired membrane, dead and viable cells were labelled with Annexin V and Propidium Iodide after 24h cold storage, followed by AFM measurement and Young's modulus of cells was derived. Viable hMSCs have a Young's modulus (E) in the range of 0.81-1.13kPa and consistent measurement was observed when different measurement locations were chosen. E of cells with partially impaired membrane was 0.69±0.17kPa or in the range of 2.04-4.74kPa, depending upon the measurement locations. With the loss of membrane integrity, though there was no variation on measured E between different locations, a mixed picture of cell stiffness was observed as indicated by cells with E as low as 0.09±0.03kPa, in a mid-range of 4.62±0.67kPa, and the highest of up to 48.98±19.80kPa. With the progress of cell death, the highest stiffness was noticed for cells showing a more granular appearance; also the lowest stiffness for cells with vacuole appearance. Findings from this study indicate that cell stiffness is significantly altered with the progress of cell death.
本研究通过原子力显微镜(AFM)观察到,在经过 24 小时的冷存后,人类间充质干细胞(hMSC)的膜损伤、死亡和存活细胞分别用 Annexin V 和碘化丙啶进行标记,随后进行 AFM 测量,得出细胞的杨氏模量。活的 hMSC 的杨氏模量(E)在 0.81-1.13kPa 范围内,当选择不同的测量位置时,观察到一致的测量结果。部分膜损伤的细胞的 E 值为 0.69±0.17kPa 或 2.04-4.74kPa,这取决于测量位置。随着膜完整性的丧失,尽管不同位置的测量 E 值没有变化,但观察到细胞硬度的混合图像,表现为 E 值低至 0.09±0.03kPa、中范围为 4.62±0.67kPa 以及高达 48.98±19.80kPa 的细胞。随着细胞死亡的进展,出现更颗粒状外观的细胞表现出最高的硬度;而出现空泡外观的细胞表现出最低的硬度。本研究的结果表明,细胞硬度随着细胞死亡的进展而显著改变。