| Isolation of Murine Adipose Tissue-derived Mesenchymal Stromal Cells (mASCs) and the Analysis of Their Proliferation in vitro
|
|
Author:
Date:
2015-11-05
[Abstract] Mesenchymal stromal cells (MSCs) are non-hematopoietic, perivascular cells which support hematopoiesis and are thought to participate in tissue repair in vivo. MSCs can be isolated from various tissues and organs and are defined in vitro as plastic adherent cells expressing CD73, CD90, CD105 (human MSCs) or CD29, CD44, sca-1 (murine MSCs) which can differentiate into osteoblasts, adipocytes, chondroblasts and myocytes. MSCs possess potent immunomodulatory and trophic capacities in vitro and in vivo and have thus emerged as a promising treatment of inflammatory/autoimmune diseases. The use of MSCs for human disease relies on the injection of a large number of cells and much effort has been focused on acquiring MSCs with high proliferative capacity. Thus, ...
[摘要] 间充质基质细胞（MSC）是非造血的血管周围细胞，其支持造血并且被认为在体内参与组织修复。 MSC可以从各种组织和器官中分离并在体外定义为表达CD73，CD90，CD105（人MSC）或CD29，CD44，sca-1（鼠MSC）的塑性贴壁细胞，其可以分化成骨细胞，脂肪细胞，软骨细胞和肌细胞。 MSC在体外具有强的免疫调节和营养能力，并且在体内具有强的免疫调节能力，因此已经成为炎症/自身免疫疾病的有希望的治疗。 MSC对人类疾病的使用依赖于大量细胞的注射，并且许多努力已经集中在获得具有高增殖能力的MSC。因此，建立简单和准确的测定MSC增殖的协议对于基础研究和应用研究都是重要的。目前的方案提供了关于如何使用xCELLigence系统和CellTiter-Blue试剂（Carrillo-Galvez等人，）分离和测量源自腹股沟和/或腹内脂肪组织（mASCs）的鼠MSC的增殖的细节>，2015; Anderson ，，2013）。下面描述的方案也可以容易地翻译成人MSC。
|
|
|
| siRNA Screening for Genes Involved in HSV-1 Replication
|
|
Author:
Date:
2014-08-20
[Abstract] Small interfering RNAs (siRNAs) are small (typically 18-24 nucleotides) RNA molecules capable of silencing gene expression post-transcriptionally and as such, they provide a simple method by which the role of individual genes in complex cellular systems can be easily assessed. As siRNAs are easy to use experimentally, and can be designed to target any gene (including pathogens), their use is perfectly suited to and easily adapted to high-throughput genome-wide screening methodologies and a range of phenotypic assays. Here we describe the use of a large siRNA library (>8,000 genes targeted individually) to screen for and identify host factors functionally involved in the replication of a human herpesvirus (Herpes simplex virus type 1; HSV-1) (Griffiths et al., 2013; Griffiths, ...
[摘要] 小干扰RNA（siRNA）是能够沉默转录后基因表达的小（通常为18-24个核苷酸）RNA分子，因此，它们提供了一种简单的方法，通过其可以容易地评估单个基因在复杂细胞系统中的作用。 由于siRNA易于实验使用，并且可以设计为靶向任何基因（包括病原体），它们的使用完全适合于并且容易地适应于高通量全基因组筛选方法学和一系列表型测定。 这里我们描述了使用大的siRNA文库（> 8,000个基因单独靶向）来筛选和鉴定功能上涉及人疱疹病毒（1型单纯疱疹病毒; HSV-1）（Griffiths）的复制的宿主因子， et al。，2013; Griffiths，2013）。
|
|
|