| Histological and Immunohistochemical Examination of Stem Cell Proliferation and Reepithelialization in the Wounded Skin
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Author:
Date:
2021-01-20
[Abstract] The skin is the largest organ that protects our body from the external environment and it is constantly exposed to pathogenic insults and injury. Repair of damage to this organ is carried out by a complex process involving three overlapping phases of inflammation, proliferation and remodeling. Histological analysis of wounded skin is a convenient approach to examine broad alterations in tissue architecture and investigate cells in their indigenous microenvironment. In this article we present a protocol for immunohistochemical examination of wounded skin to study mechanisms involved in regulating stem cell activity, which is a vital component in the repair of the damaged tissue. Performing such histological analysis enables the understanding of the spatial relationship between cells that ...
[摘要] [摘要]皮肤是保护我们的身体免受外界环境影响最大的器官,并且不断受到病原体的伤害和伤害。修复该器官的水坝年龄是通过复杂的过程进行的,该过程涉及炎症,增殖和重塑的三个重叠阶段。对受伤的皮肤进行组织学分析是检查组织结构的广泛变化并研究其固有微环境中的细胞的便捷方法。在本文中,我们提出了一种用于受伤皮肤的免疫组织化学检查的方案,以研究参与调节干细胞活性的机制,这是修复受损组织的重要组成部分。进行这种组织学分析可以了解在特殊伤口微环境中相互作用的细胞之间的空间关系。本文所述的分析工具,允许邻近伤口和重新程度的干细胞的再生能力的定量测量-伤口闭合过程中的上皮。这些协议可以适用于研究受伤皮肤内的众多细胞过程和细胞类型。
[背景]皮肤具有巨大的再生能力,这归因于位于整个器官不同部位的众多常驻干细胞种群的存在。免疫组织学方法与目标细胞的遗传标记相结合,已大大提高了体内对干细胞调节和组织稳态的认识(Blanpain等,2004)。这些方法已用于鉴定与毛囊相关的多个干细胞群体,包括位于毛发隆突中的表皮和黑素细胞干细胞,次级毛发芽中的快速循环祖细胞以及真皮鞘中存在的真皮干细胞(Greco等。,2009;拉赫马尼等人,2020)。这些干细胞具有自我更新,多能性和高度增殖性,可在生理性毛发循环过程中再生组织,并在伤口愈合过程中修复损伤。引人注目的是,由于这些细胞分裂并迅速向伤口部位进展,使上皮中的干细胞对皮肤损伤迅速做出反应,从而使伤口重新上皮化(Ito等人,2005)。 ...
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| Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis
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Author:
Date:
2021-01-05
[Abstract] The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain slices are emerging as a valuable tool. Over the recent years, this methodology has been refined and here we present a combined protocol that brings together these refinements to enable long-term culture of adult hippocampal slices. We employ a sectioning technique that retains essential afferent inputs onto the hippocampus as well as serum-free culture conditions, so allowing an extended culture period. To sustain the neurogenic potential in the ...
[摘要] [摘要]海马的功能取决于成年海马神经发生的过程,该过程是这种结构异常的神经发育的基础,并且在中枢神经系统病理中也经常受到影响。因此,对该过程的操纵代表了重要的治疗目标。为了确定潜在的策略,器官型成人大脑切片正在成为一种有价值的工具。近年来,此方法已得到完善,在此我们提出一种组合协议, 汇集了这些改进,以实现成人海马切片的长期培养。我们采用了一种切片技术,可将必要的传入输入保留在海马上以及无血清培养条件下,因此可以延长培养时间。为了维持切片中的神经源性潜力,我们利用神经胶质生成抑制剂吲哚美辛。使用EdU保留分析使我们能够评估药理干预对神经发生的影响。通过这些改进,我们建立了一种简单可靠的方法来研究小分子/药物对离体增殖和神经元形成的影响,这将有助于未来发现驱动的药物筛选。
[背景技术]海马是具有高度的可塑性作为整个生命齿状回中正在进行的神经发生的结果,脑的独特区域。成年海马神经发生的这一过程始于在亚颗粒区(SGZ)中神经干细胞(NSC)的不对称分裂,该过程保留了干细胞池并生成了准备用于神经元分化的祖细胞(Kempermann等人,2004;Anacker和Hen ,2017; ...
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