Department of Paediatrics and Child Neurology, Emma Children's Hospital, Amsterdam University Medical Centre, Meibergdreef 9, Amsterdam, 1100 DD, The Netherlands.
Amsterdam Leukodystrophy Center, Emma Children's Hospital, Amsterdam University Medical Centre, Amsterdam Neuroscience, Cellular & Molecular Mechanisms, Meibergdreef 9, 1100 DD, Amsterdam, The Netherlands.
Acta Neuropathol Commun. 2024 May 31;12(1):83. doi: 10.1186/s40478-024-01784-1.
Human brain experimental models recapitulating age- and disease-related characteristics are lacking. There is urgent need for human-specific tools that model the complex molecular and cellular interplay between different cell types to assess underlying disease mechanisms and test therapies. Here we present an adapted ex vivo organotypic slice culture method using human post-mortem brain tissue cultured at an air-liquid interface to also study brain white matter. We assessed whether these human post-mortem brain slices recapitulate the in vivo neuropathology and if they are suitable for pathophysiological, experimental and pre-clinical treatment development purposes, specifically regarding leukodystrophies. Human post-mortem brain tissue and cerebrospinal fluid were obtained from control, psychiatric and leukodystrophy donors. Slices were cultured up to six weeks, in culture medium with or without human cerebrospinal fluid. Human post-mortem organotypic brain slice cultures remained viable for at least six weeks ex vivo and maintained tissue structure and diversity of (neural) cell types. Supplementation with cerebrospinal fluid could improve slice recovery. Patient-derived organotypic slice cultures recapitulated and maintained known in vivo neuropathology. The cultures also showed physiologic multicellular responses to lysolecithin-induced demyelination ex vivo, indicating their suitability to study intrinsic repair mechanisms upon injury. The slice cultures were applicable for various experimental studies, as multi-electrode neuronal recordings. Finally, the cultures showed successful cell-type dependent transduction with gene therapy vectors. These human post-mortem organotypic brain slice cultures represent an adapted ex vivo model suitable for multifaceted studies of brain disease mechanisms, boosting translation from human ex vivo to in vivo. This model also allows for assessing potential treatment options, including gene therapy applications. Human post-mortem brain slice cultures are thus a valuable tool in preclinical research to study the pathomechanisms of a wide variety of brain diseases in living human tissue.
目前缺乏能够重现年龄相关性和疾病相关性特征的人类大脑实验模型。因此，我们迫切需要人类特异性工具，以模拟不同细胞类型之间复杂的分子和细胞相互作用，从而评估潜在的疾病机制并测试治疗方法。在这里，我们介绍了一种经过改良的离体器官型切片培养方法，使用在气液界面培养的人类死后大脑组织来研究大脑白质。我们评估了这些人类死后大脑切片是否能够重现体内神经病理学特征，以及它们是否适合用于生理病理学、实验和临床前治疗开发目的，特别是针对脑白质营养不良。我们从对照、精神科和脑白质营养不良供体中获得了人类死后脑组织和脑脊液。将切片在含有或不含有人类脑脊液的培养基中培养长达 6 周。人类死后器官型大脑切片培养物在体外至少能存活 6 周，并且保持组织结构和（神经）细胞类型的多样性。补充脑脊液可以改善切片的恢复。患者来源的器官型切片培养物重现和维持了已知的体内神经病理学特征。这些培养物还表现出对溶卵磷脂诱导的脱髓鞘的生理性多细胞反应，表明它们适合在体外研究损伤后的内在修复机制。这些切片培养物适用于各种实验研究，例如多电极神经元记录。最后，该培养物显示出成功的、依赖细胞类型的基因治疗载体转导。这些人类死后器官型大脑切片培养物代表了一种适合多种大脑疾病机制研究的适应性体外模型，促进了从人类体外到体内的转化。该模型还允许评估潜在的治疗选择，包括基因治疗应用。因此，人类死后大脑切片培养物是临床前研究中的一种有价值的工具，可用于在活体人类组织中研究各种脑疾病的病理机制。
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