Institute of Biomedicine, University of Eastern Finland, 70211 Kuopio, Finland.
A.I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, 70211 Kuopio, Finland.
Cells. 2023 May 18;12(10):1422. doi: 10.3390/cells12101422.
Organotypic slice culture models surpass conventional in vitro methods in many aspects. They retain all tissue-resident cell types and tissue hierarchy. For studying multifactorial neurodegenerative diseases such as tauopathies, it is crucial to maintain cellular crosstalk in an accessible model system. Organotypic slice cultures from postnatal tissue are an established research tool, but adult tissue-originating systems are missing, yet necessary, as young tissue-originating systems cannot fully model adult or senescent brains. To establish an adult-originating slice culture system for tauopathy studies, we made hippocampal slice cultures from transgenic 5-month-old hTau.P301S mice. In addition to the comprehensive characterization, we set out to test a novel antibody for hyperphosphorylated TAU (pTAU, B6), with and without a nanomaterial conjugate. Adult hippocampal slices retained intact hippocampal layers, astrocytes, and functional microglia during culturing. The P301S-slice neurons expressed pTAU throughout the granular cell layer and secreted pTAU to the culture medium, whereas the wildtype slices did not. Additionally, cytotoxicity and inflammation-related determinants were increased in the P301S slices. Using fluorescence microscopy, we showed target engagement of the B6 antibody to pTAU-expressing neurons and a subtle but consistent decrease in intracellular pTAU with the B6 treatment. Collectively, this tauopathy slice culture model enables measuring the extracellular and intracellular effects of different mechanistic or therapeutic manipulations on TAU pathology in adult tissue without the hindrance of the blood-brain barrier.
器官型切片培养模型在许多方面优于传统的体外方法。它们保留了所有组织驻留的细胞类型和组织层次结构。对于研究多因素神经退行性疾病,如 tau 病,在可访问的模型系统中保持细胞串扰至关重要。来自新生组织的器官型切片培养物是一种成熟的研究工具,但缺乏源自成年组织的系统,然而这是必要的,因为年轻组织来源的系统不能完全模拟成年或衰老的大脑。为了建立用于 tau 病研究的成年起源切片培养系统,我们从转基因 5 个月大的 hTau.P301S 小鼠中制作了海马切片培养物。除了全面表征外,我们还着手测试一种用于磷酸化 tau (pTAU, B6) 的新型抗体,包括有无纳米材料缀合物。成年海马切片在培养过程中保留了完整的海马层、星形胶质细胞和功能性小胶质细胞。P301S 切片神经元在颗粒细胞层中表达 pTAU 并将其分泌到培养基中,而野生型切片则没有。此外,P301S 切片中的细胞毒性和炎症相关决定因素增加。使用荧光显微镜,我们显示了 B6 抗体与表达 pTAU 的神经元的靶标结合,以及 B6 处理后细胞内 pTAU 的细微但一致减少。总的来说,这种 tau 病切片培养模型能够测量不同机制或治疗干预对成年组织中 TAU 病理学的细胞外和细胞内影响,而不受血脑屏障的阻碍。