Optimization of long-term human iPSC-derived spinal motor neuron culture using a dendritic polyglycerol amine-based substrate
| Autor: | Stefano Stifani, Jean-Pierre Clément, Louise Thiry, Timothy E. Kennedy, Rainer Haag |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Glycerol
Cell type Polymers Human iPSCs Induced Pluripotent Stem Cells Neurosciences. Biological psychiatry. Neuropsychiatry Context (language use) multi-electrode array Biology single cell RNA sequencing medicine Humans spinal motor neurons Viability assay Amines Amyotrophic lateral sclerosis Motor Neurons Experimental model General Neuroscience Cell Differentiation 500 Naturwissenschaften und Mathematik::570 Biowissenschaften Biologie::570 Biowissenschaften Biologie Spinal muscular atrophy Motor neuron medicine.disease Original Papers Electrophysiology medicine.anatomical_structure dendritic polyglycerol amine Neurology (clinical) Neuroscience RC321-571 |
| Zdroj: | ASN Neuro, Vol 14 (2022) ASN NEURO |
| DOI: | 10.1101/2021.09.14.460098 |
| Popis: | Human induced pluripotent stem cells (hiPSCs) derived from healthy and diseased individuals can give rise to many cell types, facilitating the study of mechanisms of development, human disease modeling, and early drug target validation. In this context, experimental model systems based on hiPSC-derived motor neurons (MNs) have been used to study MN diseases such as spinal muscular atrophy and amyotrophic lateral sclerosis. Modeling MN disease using hiPSC-based approaches requires culture conditions that can recapitulate in a dish the events underlying differentiation, maturation, aging, and death of MNs. Current hiPSC-derived MN-based applications are often hampered by limitations in our ability to monitor MN morphology, survival, and other functional properties over a prolonged timeframe, underscoring the need for improved long-term culture conditions. Here we describe a cytocompatible dendritic polyglycerol amine (dPGA) substrate-based method for prolonged culture of hiPSC-derived MNs. We provide evidence that MNs cultured on dPGA-coated dishes are more amenable to long-term study of cell viability, molecular identity, and spontaneous network electrophysiological activity. The present study has the potential to improve hiPSC-based studies of human MN biology and disease.We describe the use of a new coating substrate providing improved conditions for long-term cultures of human iPSC-derived motor neurons, thus allowing evaluation of cell viability, molecular identity, spontaneous network electrophysiological activity, and single-cell RNA sequencing of mature motor neurons. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|