Benwood Claire, Walters-Shumka Jonathan, Scheck Kali, Willerth Stephanie M
Department of Mechanical Engineering, University of Victoria, Victoria, BC, V8P 5C2, Canada.
Division of Medical Sciences, University of Victoria, Victoria, BC, V8P 5C2, Canada.
Bioelectron Med. 2023 May 24;9(1):10. doi: 10.1186/s42234-023-00112-7.
Alzheimer's disease (AD), a progressive neurodegenerative disorder, is becoming increasingly prevalent as our population ages. It is characterized by the buildup of amyloid beta plaques and neurofibrillary tangles containing hyperphosphorylated-tau. The current treatments for AD do not prevent the long-term progression of the disease and pre-clinical models often do not accurately represent its complexity. Bioprinting combines cells and biomaterials to create 3D structures that replicate the native tissue environment and can be used as a tool in disease modeling or drug screening.
This work differentiated both healthy and diseased patient-derived human induced pluripotent stems cells (hiPSCs) into neural progenitor cells (NPCs) that were bioprinted using the Aspect RX1 microfluidic printer into dome-shaped constructs. The combination of cells, bioink, and puromorphamine (puro)-releasing microspheres were used to mimic the in vivo environment and direct the differentiation of the NPCs into basal forebrain-resembling cholinergic neurons (BFCN). These tissue models were then characterized for cell viability, immunocytochemistry, and electrophysiology to evaluate their functionality and physiology for use as disease-specific neural models.
Tissue models were successfully bioprinted and the cells were viable for analysis after 30- and 45-day cultures. The neuronal and cholinergic markers β-tubulin III (Tuj1), forkhead box G1 (FOXG1), and choline acetyltransferase (ChAT) were identified as well as the AD markers amyloid beta and tau. Further, immature electrical activity was observed when the cells were excited with potassium chloride and acetylcholine.
This work shows the successful development of bioprinted tissue models incorporating patient derived hiPSCs. Such models can potentially be used as a tool to screen promising drug candidates for treating AD. Further, this model could be used to increase the understanding of AD progression. The use of patient derived cells also shows the potential of this model for use in personalized medicine applications.
阿尔茨海默病（AD）是一种进行性神经退行性疾病，随着人口老龄化，其发病率日益上升。其特征是β淀粉样蛋白斑块的积累以及含有高度磷酸化tau的神经原纤维缠结。目前针对AD的治疗方法无法阻止疾病的长期进展，临床前模型往往不能准确反映其复杂性。生物打印将细胞和生物材料结合起来，创建三维结构，以复制天然组织环境，并可用作疾病建模或药物筛选的工具。
这项工作将健康和患病患者来源的人诱导多能干细胞（hiPSC）分化为神经祖细胞（NPC），使用Aspect RX1微流控打印机将其生物打印成圆顶形构建体。细胞、生物墨水和释放嘌呤霉素（puro）的微球组合用于模拟体内环境，并引导NPC分化为类似基底前脑的胆碱能神经元（BFCN）。然后对这些组织模型进行细胞活力、免疫细胞化学和电生理学表征，以评估其作为疾病特异性神经模型的功能和生理学特性。
成功地对组织模型进行了生物打印，细胞在培养30天和45天后仍具有活性可供分析。鉴定出神经元和胆碱能标记物β-微管蛋白III（Tuj1）、叉头框G1（FOXG1）和胆碱乙酰转移酶（ChAT）以及AD标记物β淀粉样蛋白和tau。此外，当用氯化钾和乙酰胆碱刺激细胞时，观察到了未成熟的电活动。
这项工作表明成功开发了包含患者来源hiPSC的生物打印组织模型。此类模型有可能用作筛选治疗AD的有前景候选药物的工具。此外，该模型可用于增进对AD进展的理解。使用患者来源的细胞也显示了该模型在个性化医疗应用中的潜力。
Zotero 插件
