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Practical guide for preparation, computational reconstruction and analysis of 3D human neuronal networks in control and ischaemic conditions.
Räsänen, Noora; Harju, Venla; Joki, Tiina; Narkilahti, Susanna.
Afiliación
  • Räsänen N; Faculty of Medicine and Health Technology, Tampere University, 33100, Tampere, Finland.
  • Harju V; Faculty of Medicine and Health Technology, Tampere University, 33100, Tampere, Finland.
  • Joki T; Faculty of Medicine and Health Technology, Tampere University, 33100, Tampere, Finland.
  • Narkilahti S; Faculty of Medicine and Health Technology, Tampere University, 33100, Tampere, Finland.
Development ; 149(20)2022 10 15.
Article en En | MEDLINE | ID: mdl-35929583
To obtain commensurate numerical data of neuronal network morphology in vitro, network analysis needs to follow consistent guidelines. Important factors in successful analysis are sample uniformity, suitability of the analysis method for extracting relevant data and the use of established metrics. However, for the analysis of 3D neuronal cultures, there is little coherence in the analysis methods and metrics used in different studies. Here, we present a framework for the analysis of neuronal networks in 3D. First, we selected a hydrogel that supported the growth of human pluripotent stem cell-derived cortical neurons. Second, we tested and compared two software programs for tracing multi-neuron images in three dimensions and optimized a workflow for neuronal analysis using software that was considered highly suitable for this purpose. Third, as a proof of concept, we exposed 3D neuronal networks to oxygen-glucose deprivation- and ionomycin-induced damage and showed morphological differences between the damaged networks and control samples utilizing the proposed analysis workflow. With the optimized workflow, we present a protocol for preparing, challenging, imaging and analysing 3D human neuronal cultures.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Madre Pluripotentes / Neuronas Límite: Humans Idioma: En Revista: Development Asunto de la revista: BIOLOGIA / EMBRIOLOGIA Año: 2022 Tipo del documento: Article País de afiliación: Finlandia Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Madre Pluripotentes / Neuronas Límite: Humans Idioma: En Revista: Development Asunto de la revista: BIOLOGIA / EMBRIOLOGIA Año: 2022 Tipo del documento: Article País de afiliación: Finlandia Pais de publicación: Reino Unido