South Australian Health and Medical Research Institute (SAHMRI), Laboratory for Human Neurophysiology and Genetics, Adelaide, SA, Australia; Flinders University, Flinders Health and Medical Research Institute (FHMRI), College of Medicine and Public Health, Adelaide, SA, Australia.
TekCyte Limited, Adelaide, SA, Australia; Cooperative Research Centre for Cell Therapy Manufacturing (CTM CRC), Adelaide, SA, Australia.
Stem Cell Reports. 2022 Mar 8;17(3):489-506. doi: 10.1016/j.stemcr.2022.01.013. Epub 2022 Feb 17.
Advances in cellular reprogramming have radically increased the use of patient-derived cells for neurological research in vitro. However, adherence of human neurons on tissue cultureware is unreliable over the extended periods required for electrophysiological maturation. Adherence issues are particularly prominent for transferable glass coverslips, hindering imaging and electrophysiological assays. Here, we assessed thin-film plasma polymer treatments, polymeric factors, and extracellular matrix coatings for extending the adherence of human neuronal cultures on glass. We find that positive-charged, amine-based plasma polymers improve the adherence of a range of human brain cells. Diaminopropane (DAP) treatment with laminin-based coating optimally supports long-term maturation of fundamental ion channel properties and synaptic activity of human neurons. As proof of concept, we demonstrated that DAP-treated glass is ideal for live imaging, patch-clamping, and optogenetics. A DAP-treated glass surface reduces the technical variability of human neuronal models and enhances electrophysiological maturation, allowing more reliable discoveries of treatments for neurological and psychiatric disorders.
细胞重编程技术的进步极大地增加了患者来源的细胞在体外神经科学研究中的应用。然而，在电生理成熟所需的延长时间内，人类神经元在组织培养器皿上的黏附并不可靠。对于可转移的玻璃盖玻片，黏附问题尤为突出，这阻碍了成像和电生理测定。在这里，我们评估了薄膜等离子体聚合物处理、聚合物因素和细胞外基质涂层，以延长人类神经元在玻璃上的黏附。我们发现带正电荷的、基于氨基的等离子体聚合物可以改善一系列人脑细胞的黏附。用层粘连蛋白涂层处理二氨基丙烷（DAP）可最佳支持人类神经元基本离子通道特性和突触活性的长期成熟。作为概念验证，我们证明了 DAP 处理的玻璃非常适合活细胞成像、膜片钳和光遗传学。DAP 处理的玻璃表面减少了人类神经元模型的技术变异性，并增强了电生理成熟，从而更可靠地发现治疗神经和精神疾病的方法。
