PDF(Pubmed)
Abstract：
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.
摘要：
缺乏概括年龄和疾病相关特征的人脑实验模型。迫切需要模拟不同细胞类型之间复杂的分子和细胞相互作用的人类特异性工具，以评估潜在的疾病机制和测试疗法。在这里，我们提出了一种适应的离体器官切片培养方法，该方法使用在气液界面上培养的人类死后脑组织来研究脑白质。我们评估了这些人类死后脑切片是否概括了体内神经病理学，以及它们是否适合于病理生理学，实验和临床前治疗的发展目的，特别是关于脑白质营养不良。从对照中获得人类死后脑组织和脑脊液，精神病和脑白质营养不良的捐献者.切片培养长达六周，在有或没有人脑脊液的培养基中。人类死后器官型脑切片培养物至少在体外保持存活六周，并保持了组织结构和（神经）细胞类型的多样性。补充脑脊液可以改善切片恢复。患者来源的器官型切片培养物概括并维持已知的体内神经病理学。培养物还显示了对溶血卵磷脂诱导的离体脱髓鞘的生理性多细胞反应，表明它们适合研究损伤后的内在修复机制。切片培养物适用于各种实验研究，作为多电极神经元记录。最后,培养物显示基因治疗载体成功的细胞类型依赖性转导.这些人类死后器官型脑切片培养物代表了一种适应的离体模型，适用于脑部疾病机制的多方面研究。促进从人类离体到体内的翻译。该模型还允许评估潜在的治疗方案，包括基因治疗应用。因此，人类死后脑切片培养是临床前研究中研究活体人体组织中各种脑疾病的病理机制的宝贵工具。
