Characterization of putative stem cell populations in the cornea using synchrotron infrared microspectroscopy
Characterization of putative stem cell populations in the cornea using synchrotron infrared microspectroscopy
复制标题
DOI：
10.1167/iovs.05-1254
复制
发表时间：
2006-06-01
影响因子：
4.4
通讯作者：
Fullwood, Nigel J.
中科院分区：
文献类型：
--
作者：
German, Matthew J.;Pollock, Hubert M.;Fullwood, Nigel J.
PUROSE。采用高分辨率同步辐射傅里叶变换红外（SR-FTIR）显微光谱技术结合多变量分析，研究了角膜上皮中成体干细胞（SC）、瞬时扩增（TA）细胞和终末分化（TD）细胞群的特征。通过使用达雷斯伯里实验室（英国）的同步加速器显微光谱设备收集牛角膜冷冻切片中原位单个细胞的光谱。通过多变量分析来分析所得光谱。三种不同细胞群的中值光谱显示出显著差异，这与它们的分化程度和增殖能力相关。多变量（主成分）分析（PCA）表明，这三个细胞群可以分离成离散的集群，只有轻微的重叠之间的SC和TA细胞群。进一步分析（Mann-Whitney检验）表明SC和TA细胞群之间最显著（P < 0.001）的光谱差异主要与核酸构象的变化有关。SR-FTIR显微光谱加上PCA似乎能够识别SC，TA细胞和TD细胞群。结果还表明，角膜上皮SC龛中的一小部分细胞亚群具有TA细胞样特征。与SC相关的最显著的光谱特征似乎与核酸构象的差异有关。这一发现与最近的理论一致，表明分化的控制与染色质结构的重大变化有关。
PUROSE. High-resolution synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy coupled with multivariate analysis was used to investigate the characteristics of putative adult Stem cell (SC), transiently amplified (TA) cell, and terminally differentiated (TD) cell populations of the corneal epithelium.METHODS. Spectra of individual cells in situ in cryosections of bovine cornea were collected by using a synchrotron microspectroscopy facility at Daresbury Laboratory (United Kingdom). The resultant spectra were analyzed by multivariate analysis.RESULTS. The median spectra of the three different cell populations showed marked differences, which correlated with their degree of differentiation and proliferative capacity. Multivariate (principal component) analysis (PCA) showed that the three cell populations could be segregated into discrete clusters, with only a slight overlap between the SC and TA cell populations. Further analysis (Mann-Whitney test) indicated that the most significant (P < 0.001) spectral differences between the SC and TA cell populations were chiefly associated with changes in nucleic acid conformation.CONCLUSIONS. SR-FTIR microspectroscopy coupled with PCA appears to enable the identification of SC, TA cell, and TD cell populations. The results also suggest that a small subpopulation of cells in the corneal epithelial SC niche possess TA cell-like characteristics. The most significant spectral characteristics associated with the SCs appear to relate to differences in nucleic acid conformation. This finding is consistent with recent theories suggesting that the control of differentiation is related to major changes in chromatin structure.