Lundin Brady F, Knight Gavin T, Fedorchak Nikolai J, Krucki Kevin, Iyer Nisha, Maher Jack E, Izban Nicholas R, Roberts Abilene, Cicero Madeline R, Robinson Joshua F, Iskandar Bermans J, Willett Rebecca, Ashton Randolph S
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53705, USA.
Medical Scientist Training Program, University of Wisconsin-Madison, School of Medicine and Public Health, Madison, WI, 53705 USA.
bioRxiv. 2024 Jun 20:2024.04.01.587605. doi: 10.1101/2024.04.01.587605.
Neural organoids have revolutionized how human neurodevelopmental disorders (NDDs) are studied. Yet, their utility for screening complex NDD etiologies and in drug discovery is limited by a lack of scalable and quantifiable derivation formats. Here, we describe the RosetteArray platform's ability to be used as an off-the-shelf, 96-well plate assay that standardizes incipient forebrain and spinal cord organoid morphogenesis as micropatterned, 3-D, singularly polarized neural rosette tissues (>9000 per plate). RosetteArrays are seeded from cryopreserved human pluripotent stem cells, cultured over 6-8 days, and immunostained images can be quantified using artificial intelligence-based software. We demonstrate the platform's suitability for screening developmental neurotoxicity and genetic and environmental factors known to cause neural tube defect risk. Given the presence of rosette morphogenesis perturbation in neural organoid models of NDDs and neurodegenerative disorders, the RosetteArray platform could enable quantitative high-throughput screening (qHTS) of human neurodevelopmental risk across regulatory and precision medicine applications.
神经类器官彻底改变了人类神经发育障碍（NDDs）的研究方式。然而，由于缺乏可扩展和可量化的衍生形式，它们在筛查复杂的NDD病因及药物发现方面的效用受到限制。在此，我们描述了玫瑰花结阵列平台用作现成的96孔板检测方法的能力，该方法将早期前脑和脊髓类器官形态发生标准化为微图案化的三维单极化神经玫瑰花结组织（每板>9000个）。玫瑰花结阵列由冷冻保存的人类多能干细胞接种，培养6 - 8天，免疫染色图像可使用基于人工智能的软件进行量化。我们证明了该平台适用于筛查已知会导致神经管缺陷风险的发育性神经毒性以及遗传和环境因素。鉴于在NDDs和神经退行性疾病的神经类器官模型中存在玫瑰花结形态发生扰动，玫瑰花结阵列平台可实现跨监管和精准医学应用的人类神经发育风险的定量高通量筛选（qHTS）。
