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DPBS, no calcium, no magnesium

DPBS,无钙,无镁

Company: Thermo Fisher Scientific
Catalog#: 14190144
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Isolation and Establishment of Mesenchymal Stem Cells from Wharton’s Jelly of Human Umbilical Cord
Author:
Date:
2018-02-20
[Abstract]  Mesenchymal stem cells (MSCs) are currently considered as ‘medicinal signaling cells’ and a promising resource in regard to cell-based regenerative therapy. Umbilical cord is a human term perinatal tissue which is easily attainable, and a promising source of stem cells with no associated ethical concerns. MSCs have been isolated from different regions of the umbilical cord and Wharton’s jelly (WJ) is the gelatinous matrix that surrounds and provides protection to the umbilical cord blood vessels. Being more primitive, MSCs from human umbilical cord exhibit greater proliferative capacity and immunosuppressive ability as compared to adult stem cells which gives them a therapeutic advantage. To meet the requirements for cell therapy, it is important to generate MSCs at a clinical scale by ... [摘要]  间充质干细胞(mesenchymal stem cells,MSCs)目前被认为是“医药信号传导细胞”,在基于细胞的再生治疗方面是一种很有前景的资源。脐带是人类的围产期组织,很容易实现,是一种有前途的干细胞来源,没有相关的伦理问题。 MSC已经从脐带的不同区域分离出来,而Wharton's果冻(WJ)是包围并提供对脐带血管的保护的凝胶状基质。更原始的是,与成人干细胞相比,来自人脐带的MSC表现出更大的增殖能力和免疫抑制能力,这使其具有治疗优势。为了满足细胞疗法的要求,通过遵循不耗时或劳动强度的步骤来产生临床规模的MSC是重要的。在此我们提出了一种简单,高效的方法,通过外植体培养方法从人脐带WJ中分离出MSC,这种方法具有重现性和成本效益。

【背景】间充质干细胞(MSC)具有显着的临床潜力来治疗各种衰弱性疾病,主要是由于其独特的免疫调节作用和再生能力(Caplan and Sorrell,2015)。它们存在于许多组织中(Hass et al。,2011),并被观察到是血管周围的体内(Caplan和Correa,2011)。来源或来源本身的小生境可能导致各种MSC类型之间的重要功能差异(Kwon等人,2016年)。虽然骨髓是研究得最充分和最好的MSCs来源,但也有一定的局限性(Liu et al。,2016)。 ...

Mouse Model of Immune Complex-mediated Vasculitis in Dorsal Skin and Assessment of the Neutrophil-mediated Tissue Damage
Author:
Date:
2017-12-20
[Abstract]  Neutrophils are the most abundant leukocytes in the blood. In the recent decades, their crucial roles in host defense, immune regulation and tissue damage have been studied in a deeper dimension. In this protocol, we described a mouse model of immune complex-mediated vasculitis in the dorsal skin induced by Arthus reaction, and the subsequent analysis of edema, hemorrhage and tissue damage due to neutrophil activation by means of Evans blue area analysis, histology, and immunofluorescence. This protocol could facilitate the investigation of cellular therapy strategy against over-activated neutrophil-mediated tissue damage. [摘要]  嗜中性粒细胞是血液中最丰富的白细胞。 近几十年来,它们在宿主防御,免疫调节和组织损伤方面的关键作用已经得到了更深入的研究。 在这个协议中,我们描述了Arthus反应诱导的背部皮肤中的免疫复合物介导的血管炎的小鼠模型,以及随后通过伊文思蓝区域分析,组织学和分析由于嗜中性粒细胞活化引起的水肿,出血和组织损伤免疫荧光。 该协议可以促进细胞治疗策略对过度活化的嗜中性粒细胞介导的组织损伤的调查。

【背景】嗜中性粒细胞构成循环白细胞的最大的进化保守部分。他们引导第一波主机防御感染或组织损伤。嗜中性粒细胞介导的细胞毒性的体外模型已被充分证实(Incani等人,1981; Dallegri等人,1984; Saffarzadeh等人,等人,2012年)。但是,为了剖析中性粒细胞介导的无菌组织损伤的复杂性,“体内”模型是必不可少的。

免疫复合物(IC)介导的血管炎是由抗原 - 抗体复合物在血管中沉积引发的疾病,其随后导致补体激活,嗜中性粒细胞募集和活化。活化的嗜中性粒细胞释放的大量活性氧和蛋白酶损害血管壁的内皮衬里并导致水肿和出血(Sindrilaru等人,2007; Goerge等人, ,2008; ...

Isolation of Mouse Cardiac Neural Crest Cells and Their Differentiation into Smooth Muscle Cells
Author:
Date:
2017-09-05
[Abstract]  Cardiac neural crest cells (CNCCs) originate at the dorsal edge of the neural tube between the otic pit and the caudal edge of the 3rd somite, and migrate into the pharyngeal arches and the heart. We have shown that fibronectin (Fn1) plays an important role in the development of the CNCC by regulating the differentiation of CNCCs into vascular smooth muscle cells around pharyngeal arch arteries (Wang and Astrof, 2016). This protocol describes the isolation of CNCCs from the neural tube and from the caudal pharyngeal arches, and the differentiation of neural crest-derived cells into smooth muscle cells. This protocol was adapted from (Newgreen and Murphy, 2000; Pfaltzgraff et al., 2012). [摘要]  心脏神经嵴细胞(CNCC)起源于神经管的背部边缘,位于第3个体节的耳穴和尾缘之间,并迁移到咽弓和心脏。 我们已经表明,纤连蛋白(Fn1)通过调节CNCCs到咽弓动脉周围的血管平滑肌细胞的分化,在CNCC的发展中起重要作用(Wang and Astrof,2016)。 该方案描述了CNCC与神经管和尾尾弓的分离,以及神经嵴衍生细胞分化成平滑肌细胞。 该方案从(Newgreen和Murphy,2000; Pfaltzgraff等人,2012)改编。
【背景】以前发表的方案描述了从神经管分离神经嵴细胞。然而,在耳孔和第三体细胞之间的神经管区域中的神经嵴细胞包括有助于许多不同细胞类型的神经嵴细胞群体;例如,迷走神经嵴细胞也来自该区域。在该方案中,我们修改了用于分离心脏神经嵴细胞的常规方法。而不是使用神经管,我们在胚胎期(E)9.5(22-25个体节期)使用尾部咽部弓形区。这是在将心脏神经嵴细胞分化为血管平滑肌细胞之前。神经嵴培养物通常含有污染性间充质细胞,通常表达平滑肌基因。为了鉴定神经嵴衍生细胞,我们从以下交叉产生的胚胎中分离出神经嵴细胞:Fn1flox / flox; ROSAmTmG / mTmG雌性小鼠×Fn1 +/-;Tfap2αIRESCre/ ...

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