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Penicillin-Streptomycin (10,000 U/mL)

青霉素 - 链霉素(10,000U / mL)

Company: Thermo Fisher Scientific
Catalog#: 15140148
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Hair Follicle Stem Cell Isolation and Expansion
Author:
Date:
2018-05-20
[Abstract]  Stem cells are widely used for numerous clinical applications including limbal stem cell deficiency. Stem cell derived from the bulge region of the hair follicle have the ability to differentiate into a variety of cell types including interfollicular epidermis, hair follicle structures, sebaceous glands and corneal epithelial cells when provided the appropriate cues. Hair follicle stem cells are being studied as a valuable source of autologous stem cells to treat disease. The protocol described below details the isolation and expansion of these cells for eventual clinical application. We used a dual-reporter mouse model to visualize both isolation and eventual differentiation of these cells in a limbal stem cell-deficient mouse model. [摘要]  干细胞被广泛用于许多临床应用,包括角膜缘干细胞缺陷。 当提供适当的提示时,源自毛囊凸出区域的干细胞具有分化成多种细胞类型的能力,包括滤泡间表皮,毛囊结构,皮脂腺和角膜上皮细胞。 正在研究毛囊干细胞作为自体干细胞治疗疾病的宝贵来源。 下面描述的方案详细描述了这些细胞的最终临床应用的分离和扩增。 我们使用双报告小鼠模型来观察这些细胞在角膜缘干细胞缺陷小鼠模型中的分离和最终分化。

【背景】干细胞被广泛用于多种翻译和临床应用。一种这样的临床应用是用于治疗角膜缘干细胞缺陷(LSCD)。当角膜缘干细胞群存在功能障碍或丧失时,LSCD发生,这对于由于先天性或获得性病理而维持健康的眼表非常重要。 LSCD的主要治疗策略是从患者健康眼睛的角膜缘活检组织培养自体上皮细胞片(Pellegrini等人,1997; Shortt等人,2007) 。这种策略的局限性在于它只适用于患有单侧LSCD的患者。那些有双侧LSCD的患者必须依靠免疫相关活体供体或尸体组织的同种异体角膜缘活检。由于全身性免疫抑制治疗的需要和供体组织的有限可用性,治疗成功率降低。一些研究小组一直在研究使用培养的口腔粘膜细胞治疗LSCD并取得了一些成功。然而,这些细胞通常不能表达角膜上皮分化标记角蛋白12(Inatomi等,2006),并且经常导致外周血管新生的发展(Nakamura等人, ,2004; ...

Murine Hair Follicle Derived Stem Cell Transplantation onto the Cornea Using a Fibrin Carrier
Author:
Date:
2018-05-20
[Abstract]  The goal of this protocol is to establish a procedure for cultivating stem cells on a fibrin carrier to allow for eventual transplantation to the eye. The ability to transfer stem cells to a patient is critical for treatment for a variety of disorders and wound repair. We took hair follicle stem cells from the vibrissae of transgenic mice expressing a dual reporter gene under the control of the Tet-on system and the keratin 12 promoter (Meyer-Blazejewska et al., 2011). A clonal growth assay was performed to enrich for stem cells. Once holoclones formed they were transferred onto a fibrin carrier and cultivated to obtain a confluent epithelial cell layer. Limbal stem cell deficient (LSCD) mice were used as the transplant recipient in order to test for successful grafting and ... [摘要]  该方案的目标是建立一种在纤维蛋白载体上培养干细胞以允许最终移植到眼睛的程序。 将干细胞转移给患者的能力对于治疗各种疾病和伤口修复至关重要。 我们在Tet-on系统和角蛋白12启动子(Meyer-Blazejewska等人,2011)的控制下从表达双报告基因的转基因小鼠的触须中取出毛囊干细胞。 进行克隆生长测定以富集干细胞。 一旦形成holoclones,将它们转移到纤维蛋白载体上并培养以获得融合上皮细胞层。 将角膜缘干细胞缺陷(LSCD)小鼠用作移植受体以测试移植成功和最终分化为角膜上皮表型。

【背景】干细胞被广泛用作治疗工具,因此递送手段是必不可少的。实际上,许多研究人员和公司正在寻找将细胞输送到人体内以优化细胞存活以及整合到宿主组织中的最佳方式。注射方法已广泛用于动物模型,但往往导致生存和整合差。目前正在使用利用生物材料和手术装置的技术。一种用于输送干细胞的技术是纤维蛋白载体。纤维蛋白凝胶是可降解的生物聚合物,其可粘附于允许细胞附着,迁移和增殖的天然组织(Ehrbar et al。,2005)。纤维蛋白凝胶具有许多优点,包括生物相容性,受控降解(Kjaergard等人,1994; Sidelmann等人,2000),均匀细胞分布和高细胞接种效率(Swartz ,2005)。已经将纤维蛋白凝胶用于治疗皮肤烧伤(Pellegrini等人,1999; ...

Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection
Author:
Date:
2018-05-05
[Abstract]  Human norovirus is the most common cause of acute gastroenteritis worldwide, resulting in estimated mortality of ~210,000 each year, of whom most are children under the age of five. However, norovirus can infect people of all age groups. There is a risk of prolonged infection in children, the elderly and patients who are immunocompromised. To study the inhibition of persistent norovirus replication by small molecule antivirals in vivo, we used a murine norovirus CR6 strain (MNV.CR6). We demonstrated earlier that efficient small molecules can reduce viral shedding in the stool of infected mice. Here we present how to generate the MNV.CR6 virus stock, infect type I and II interferon receptor knockout AG129 mice via oral gavage, administer antivirals to mice, and quantify viral ... [摘要]  人类诺瓦克病毒是全球急性胃肠炎最常见的原因,估计每年约有210,000人死亡,其中大部分是5岁以下的儿童。 然而,诺如病毒可以感染所有年龄组的人。 儿童,老年人和免疫功能低下的患者有长期感染的风险。 为研究体内小分子抗病毒剂对持续诺如病毒复制的抑制作用,我们使用鼠诺沃克病毒CR6株(MNV.CR6)。 我们之前表明,高效小分子可以减少感染小鼠粪便中的病毒脱落。 在这里,我们介绍如何产生MNV.CR6病毒,通过口服灌胃感染I型和II型干扰素受体敲除AG129小鼠,给小鼠施用抗病毒药物,并量化这些小鼠粪便中的病毒基因组拷贝。

【背景】人诺瓦克病毒是胃肠炎的重要原因。尽管大多数诺罗病毒感染是急性和自限性的,但是在具有免疫缺陷状态的患者中,尤其是在实体器官和造血干细胞移植受体,接受化疗的患者和患有AIDS的患者中,感染可能变成慢性的(Westhoff等人, / em>,2009; Green,2014; Angarone 等。,2016)。在幼儿和老年人中也观察到延长的诺如病毒感染,导致疾病持续时间增加,排便增加和病毒脱落长达47天(Murata等人,2007; ...

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