Invest Ophthalmol Vis Sci.:通过实时成像的方法来研究人类干细胞诱导的视网膜类器官的的结构特征和功能

2017-07-04 cuiguizhong MedSci原创

美国南加州大学凯克大学眼科学系罗斯基眼科研究所的Cobrinik D教授团队近日在Invest Ophthalmol Vis Sci发表了他们的一项重要工作,他们利用实时成像的方法来研究人类干细胞诱导的视网膜类器官的的结构特征和功能。

美国南加州大学凯克大学眼科学系罗斯基眼科研究所的Cobrinik D教授团队近日在Invest Ophthalmol Vis Sci发表了他们的一项重要工作,他们利用实时成像的方法来研究人类干细胞诱导的视网膜类器官的的结构特征和功能。

利用人类多能干细胞(hPSC)诱导的视网膜类器官是研究视网膜发育、病理生理特征和细胞治疗的良好模型。传统的组织学分析是将不同时间固定的多个标本进行分析,来重建发育过程。与此相反,视网膜类器官通过相同生物组织的重复分析来研究其变化,是更直接的手段。在体外培养过程中,新的活体成像技术可以用来研究视网膜类器官结构和代谢功能。他们的工作是采用活体组织成像技术来研究视网膜类器官的发育,如随光感受器分化过程中的代谢变化。

Cobrinik D教授团队通过相差显微镜,光学相干断层扫描(OCT),荧光寿命成像显微镜(FLIM)和高光谱成像(HSpec)等技术手段,研究不同发育阶段的hPSC诱导的视网膜类器官的微观解剖组织和代谢功能的变化特征。并与通过组织学染色、免疫染色和固定类器官组织的微电脑层析(micro-CT)的研究结果相比较。

他们将类器官切成薄片,使用FLIM和HSpec技术手段来检测代谢活动的变化。FLIM检测结果发现,糖酵解活性逐渐增加;HSpec检测发现,视网膜类器官外层的视黄醇和视黄酸积聚,这与感光体起源的研究相吻合。micro-CT方法可以用来研究视网膜类器官的三维结构,但不能用来研究类器官的薄片样本。

因此,他们认为,实时成像技术可以在视网膜类器官中实现实时和非破坏性成像。FLIM和HSpec可以用来快速检测层状结构和光感受器的代谢。实时成像技术可以大大帮助人们在不同的实验和细胞治疗的条件下连续研究视网膜类器官的发育。  Cobrinik D教授团队的这项工作给人们研究视网膜的发育提供了更多的技术手段。

原文出处:

Browne, A.W., et al., Structural and Functional Characterization of Human Stem-Cell-Derived Retinal Organoids by Live Imaging. Invest Ophthalmol Vis Sci, 2017. 58(9): p. 3311-3318.

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