{{'Search' | translate}}
 

TrypLETM Express Enzyme (1X), no phenol red

TrypLE TM Express Enzyme(1X),无酚红

Company: Thermo Fisher Scientific
Catalog#: 12604013
Bio-protocol()
Company-protocol()
Other protocol()

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)。 ...

Culture and Nucleofection of Postnatal Day 7 Cortical and Cerebellar Mouse Astroglial Cells
Author:
Date:
2018-02-05
[Abstract]  Lineage reprogramming of astroglial cells isolated from different brain regions leads to the generation of different neuronal subtypes. This protocol describes the isolation and culture of neocortical and cerebellar astrocytes from postnatal mice. We also present a comprehensive description of the main steps towards successful gene delivery in these cells using nucleofection. Neocortex and cerebellum astrocyte cultures obtained with these methods are suitable for the study of molecular and cellular mechanisms involved in direct cell lineage reprogramming into induced neurons (iNs). [摘要]  从不同脑区分离的星形胶质细胞的谱系重编程导致产生不同的神经元亚型。 该协议描述了来自出生后小鼠的新皮质和小脑星形胶质细胞的分离和培养。 因此,我们全面描述了使用nucleofection在这些细胞中成功进行基因传递的主要步骤。 在涉及直接细胞谱系重编程为诱导神经元(iNs)的分子和细胞机制研究中获得的新皮质和小脑星形胶质细胞培养物。

【背景】文献中广泛描述了星形胶质细胞培养(Saura,2007,Schildge等人,2013)。几种方案,主要是细胞分离的步骤数不同,产生足够纯度的星形胶质细胞培养物(高达98%)。然而,大多数方案被描述为“关注从新皮层分离星形胶质细胞并使用化学解离”(Schildge等人,2013)。在这里,我们提供了一种替代方法来产生高浓缩的星形胶质细胞单层,而不需要化学组织解离,这使得方案比以前的方法更快。此外,我们强调新皮层和小脑星形胶质细胞的分离和培养,突出了这两个细胞群体之间的一些重要差异。最后,我们描述了使用核转染在星形胶质细胞中基因递送的替代的,成本有效的方法(Chouchane等人,2017)。该技术由使用特定电压和试剂的细胞电穿孔后直接递送DNA分子组成。与逆转录病毒介导的转染相比(Heines等,2002; Berninger等) / ,2007,Heinrich等人,2012; ...

Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes
Author:
Date:
2018-01-20
[Abstract]  Primary mature hepatocytes (MHs) or their progenitor cells are candidate cell sources for cell transplantation therapy in severe liver diseases. However, stable culture of these cells or generation of equivalent cells from pluripotent stem cells has been limited. Using a cocktail of small molecules that we previously found useful in stable culture of multiple types of stem/progenitor cells, we recently established a novel method to generate bipotent liver progenitor cells, named chemically induced liver progenitors (CLiPs), from adult rat MHs. Here, we describe a detailed protocol for the induction of rat CLiPs. We first describe the method to isolate primary rat MHs and then describe how to induce CLiPs from these MHs. In addition, we describe a method to evaluate the bipotentiality of ... [摘要]  原代成熟肝细胞(MH)或其祖细胞是重症肝病中细胞移植治疗的候选细胞来源。然而,这些细胞的稳定培养或多能干细胞的等效细胞的产生受到限制。我们使用先前在多种类型的干/祖细胞稳定培养中发现有用的小分子混合物,最近建立了一种从成年大鼠MHs产生双能肝脏祖细胞(命名为化学诱导肝祖细胞(CLiPs))的新方法。在这里,我们描述了诱导大鼠CLiPs的详细方案。我们首先描述分离原代鼠MH的方法,然后描述如何从这些MH中诱导CLiPs。另外,我们描述了一种评估产生的CLiPs分化成肝细胞和胆管上皮细胞的双能性的方法。我们还介绍了如何通过长期的文化和详细的示例数据建立稳定的CLiP。可以在2周内产生初级CLiPs,并且可以在2.5-4个月内建立经历10次传代的稳定的CLiPs,批次间变异性。
【背景】对于实现肝病再生医学的新型细胞来源有着强烈的需求。目前唯一的治疗终末期肝病的方法是肝移植,但是由于供者短缺,其应用受到限制。最近,我们小组提出了一种产生能够在体外稳定地扩增的新型LPC的方法,并且可以以广泛的效率重新繁殖慢性肝炎动物模型的损伤肝脏(Katsuda等人, / ...

Comments