<br>3D 打印微孔板插件，用于活体脑类器官的长期高分辨率成像,BMC Biomedical Engineering

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3D 打印微孔板插件，用于活体脑类器官的长期高分辨率成像
BMC Biomedical Engineering Pub Date : 2021-04-01 , DOI: 10.1186/s42490-021-00049-5
Mariana Oksdath Mansilla ₁ , Camilo Salazar-Hernandez ₁ , Sally L Perrin ₁ , Kaitlin G Scheer ₁ , Gökhan Cildir ₁ , John Toubia _{1,

2} , Kristyna Sedivakova ₁ , Melinda N Tea ₁ , Sakthi Lenin ₁ , Elise Ponthier ₁ , Erica C F Yeo ₁ , Vinay Tergaonkar _{1,

3,

4} , Santosh Poonnoose _{5,

6} , Rebecca J Ormsby ₆ , Stuart M Pitson _{1,

7} , Michael P Brown _{1,

7,

8} , Lisa M Ebert _{1,

7,

8} , Guillermo A Gomez ₁

Affiliation

类器官是用于研究人类大脑发育和病理条件下的可靠模型。然而，目前的脑类器官培养方法产生的组织大小为 0.5 至 2 毫米，需要不断搅拌以实现适当的氧合。因此，培养条件不适合全脑类器官实时成像，而全脑类器官实时成像需要在生理相关时间范围（即天、周、月）内研究发育过程和疾病进展。在这里，我们设计了适用于标准 24 多孔板的 3D 打印微孔板插件，允许多个类器官在预定义和固定的 XYZ 坐标中生长。这项创新促进了整个大脑类器官的高分辨率成像，从而可以长期精确评估类器官的生长和形态，以及类器官内的细胞追踪。我们应用这项技术通过大脑类器官中的神经元祖细胞来跟踪新皮质的发育，以及患者来源的胶质母细胞瘤干细胞在健康大脑类器官中的运动。这个新的生物工程平台是一项重大进步，允许使用多模态倒置荧光显微镜对全脑类器官进行长期详细分析。

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3D-printed microplate inserts for long term high-resolution imaging of live brain organoids

Organoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months). Here we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids. This new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.

更新日期：2021-04-01

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