Non-invasive, quantitative assessment of the morphology of γ-irradiated human mesenchymal stem cells and periosteal cells using digital holographic microscopy
Non-invasive, quantitative assessment of the morphology of γ-irradiated human mesenchymal stem cells and periosteal cells using digital holographic microscopy
复制标题
DOI：
10.1080/09553002.2016.1230242
复制
发表时间：
2016-01-01
影响因子：
2.6
通讯作者：
Nakata, Koh
中科院分区：
文献类型：
--
作者：
Kawase, Tomoyuki;Okuda, Kazuhiro;Nakata, Koh
目的：为了确保用于细胞治疗的细胞的质量，我们检查了数字全息显微镜（DHM）用于细胞形态变化的非侵入性定量评估的适用性。材料和方法：以人牙槽骨骨膜细胞（PC）为参照，对脂肪组织间充质干细胞（MSC-AT）和骨髓间充质干细胞（MSC-BM）进行c-射线照射，（1和4戈伊），并使用全息显微镜在不固定的情况下定量其形态学变化。分离后，用乙醇固定，细胞数量和表面抗原的表达分别测定使用自动化细胞计数试剂盒和流式细胞仪。结果：在各种指标中，只有与细胞大小有关的指标在c照射后显着改变。BMC-AT和BMC-BM均较PC增大，对低剂量γ射线照射敏感。与PC相反，与DNA损伤修复相关的蛋白质（γ-H2 AX，p21（waf 1），p53和Rb）在MSC-AT或MSC-BM中均未显著上调或持续一周。结论：代替DNA损伤标记，我们建议DHM监测的细胞形态参数（例如细胞体积）可能是MSC质量的有用且更稳定的标记。
Purpose: To assure the quality of cells to be used in cell therapy, we examined the applicability of digital holographic microscopy (DHM) for non-invasive, quantitative assessment of changes in cell morphology.Materials and methods: Mesenchymal stem cells derived from adipose tissue (MSC-AT) and bone marrow (MSC-BM), in addition to human alveolar periosteal cells (PC) as a reference, were c-ray irradiated (1 and 4 Gy), and their morphological changes were quantified without fixation using holographic microscopy. After detachment and fixation with ethanol, cell number and surface antigen expression were determined using an automated cell counter kit and flow-cytometry, respectively.Results: Among various indexes, only indexes related to cell size were significantly changed after c-irradiation. Both BMC-AT and BMC-BM were enlarged and more sensitive to a low dose of gamma-irradiation than PC. In contrast to PC, proteins related to DNA damage repair (gamma-H2AX, p21(waf1), p53 and Rb) were not substantially upregulated or sustained for a week in either MSC-AT or MSC-BM.Conclusion: Instead of DNA damage markers, we suggest that cell morphological parameters (e.g. cell volume) that are monitored by DHM could be a useful and more stable marker of MSC quality.