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Human collagen IV

Company: Sigma
Catalog#: C5533
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In vitro Measurement of Membrane Attack Complex in RPE Cells
Author:
Date:
2021-02-20
[Abstract]  

Initiation of the complement system results in the formation of a multiprotein pore termed the membrane attack complex (MAC, C5b-C9). MAC pores accumulate on a cell surface and can result in cell lysis. The retinal pigment epithelium (RPE) is a single monolayer of pigmented epithelial cells located at the posterior poll of the eye that forms the outer blood retinal barrier. RPE cells are highly polarized with apical microvilli and basolateral contact with Bruch’s membrane. In order to obtain biologically relevant polarized RPE cultures in vitro, RPE cells are seeded onto the apical side of a transwell filter and cultured for 4 weeks in low serum media. MAC formation on RPE cells has been reported to be sub-lytic. MAC formation can be achieved in vitro by introduction of normal human

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[摘要]  [摘要]补体系统的启动导致形成称为膜攻击复合物(MAC,C5b-C9)的多蛋白孔。MAC孔积聚在细胞表面,可导致细胞裂解。视网膜色素上皮细胞(RPE)是位于眼那种形式的后轮询色素上皮细胞的单个单层š外血视网膜屏障。RPE细胞高度极化,顶端微绒毛和与Bruch膜的基底外侧接触。为了在体外获得生物学上相关的极化RPE培养物,将RPE细胞接种到Transwell滤膜的顶端,并在低血清培养基中培养4周。MAC形成Ò Ñ据报道,RPE细胞是亚裂解的。通过在血清饥饿24小时后向培养基中引入正常人血清(NHS),可以在体外实现MAC的形成。NHS包含启动补体激活和MAC形成所需的所有血清补体蛋白。我们结合了体外RPE极化和补体激活,以利用共聚焦显微镜在体外可视化MAC形成,从而实现了高分辨率MAC成像。


[背景]补体系统是一种进化保守的先天免疫途径。补体激活存在三种主要的独立但重叠的途径,它们在C3转化酶,经典途径,凝集素途径和替代途径中收敛。在经典途径中,免疫复合物(抗原-抗体复合物)通过C1q亚成分结合C1,然后C1s蛋白酶亚基裂解补体因子C4和C2。这些片段(C4bC2b)形成酶复合物“ ...

Intestinal Enteroid Culture for Human Astroviruses
Author:
Date:
2020-07-20
[Abstract]  Human astroviruses (HAstV) are non-enveloped, positive-sense single stranded RNA viruses that typically cause gastroenteritis in children, the elderly and among immunocompromised individuals. Some HAstV species have also been implicated in neurological diseases. It is important to study these viruses to understand the pathogenesis and develop therapeutics. Here we describe HAstV infection in epithelium-only human intestinal enteroids (HIE) isolated from biopsy-derived intestinal crypts. Although different HAstV clades have been propagated in transformed immortalized cell lines such as A549, Caco-2, HEK293T and Huh7.5, we chose HIE because they better mimic the human intestine and thus are more physiologically relevant. Additionally, HIE support the replication of all HAstV clades ... [摘要]  [摘要] 人类星状病毒(HAstV)是一种非包膜,正向单链RNA病毒,通常会导致儿童,老年人和免疫力低下的人群中的肠胃炎。一些HAstV物种也与神经系统疾病有关。研究这些病毒以了解其发病机理并开发治疗方法非常重要。在这里我们描述HAstV感染 从活检来源的肠隐窝中分离出的仅上皮的人肠小肠类固醇(HIE)。尽管已经在转化的永生化细胞系(例如A549,Caco-2,HEK293T和Huh7.5)中繁殖了不同的HAstV进化枝,但我们选择HIE是因为它们可以更好地模仿人的肠道,因此在生理上更相关。此外,HIE支持所有HAstV进化枝的复制,包括临床样品,因此使HIE成为研究HAstV生物学的有价值的潜在通用模型。

[背景] 人类星状病毒(HAstV)是高度流行的肠病毒,会引起从腹泻到脑炎的广泛感染。无症状,全身和肠道外感染很常见(Madeley和Cosgrove 1975;Bosch 等人,2014;Kolawole 等人,2019)。尽管具有医学重要性,但H AstV 是一些特征最弱的肠病毒。目前,已识别出三组人类AstV:经典HAstV和非经典HAstV-MLB(墨尔本),以及HAstV-VA / ...

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