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Bovine Serum Albumin

牛血清白蛋白

Company: Sigma-Aldrich
Catalog#: A2058
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Delivery of the Cas9 or TevCas9 System into Phaeodactylum tricornutum via Conjugation of Plasmids from a Bacterial Donor
Author:
Date:
2018-08-20
[Abstract]  Diatoms are an ecologically important group of eukaryotic microalgae with properties that make them attractive for biotechnological applications such as biofuels, foods, cosmetics and pharmaceuticals. Phaeodactylum tricornutum is a model diatom with defined culture conditions, but routine genetic manipulations are hindered by a lack of simple and robust genetic tools. One obstacle to efficient engineering of P. tricornutum is that the current selection methods for P. tricornutum transformants depend on the use of a limited number of antibiotic resistance genes. An alternative and more cost-effective selection method would be to generate auxotrophic strains of P. tricornutum by knocking out key genes involved in amino acid biosynthesis, and using ... [摘要]  硅藻是一种具有重要生态意义的真核微藻类,其特性使其对生物燃料,食品,化妆品和药品等生物技术应用具有吸引力。 Phaeodactylum tricornutum 是具有确定培养条件的模型硅藻,但缺乏简单而强大的遗传工具阻碍了常规遗传操作。有效设计 P的一个障碍。 tricornutum 是 P的当前选择方法。 tricornutum 转化体依赖于使用有限数量的抗生素抗性基因。另一种更具成本效益的选择方法是产生 P的营养缺陷型菌株。通过敲除参与氨基酸生物合成的关键基因,并使用基于质粒的生物合成基因拷贝作为选择标记,使三角酵母。以前关于 P基因敲除的研究。 tricornutum 使用biolistic转换将CRISPR-Cas9系统传递到 P.藻。非复制质粒的生物射弹转化可导致对 P的不期望的损伤。由于转化的DNA随机整合到基因组中,tricornutum 。随后固化编辑的细胞以防止Cas9的长期过表达是非常困难的,因为目前没有方法来切除整合的质粒。该协议采用新方法将Cas9或TevCas9系统传送到 P. tricornutum 通过来自细菌供体细胞的质粒的缀合。该过程涉及:1)设计和插入靶向 P的guideRNA。将tricornutum 尿素酶基因导入TevCas9表达质粒,该质粒也编码转移的接合起点,2)将该质粒安装在含有含有接合机制的质粒(pTA-Mob)的大肠杆菌中, ...

Human Endometrial Stem Cell Isolation from Endometrium and Menstrual Blood
Author:
Date:
2018-01-20
[Abstract]  Human endometrial stem cell/stromal cells (hEnSCs) are isolated from endometrium or menstrual blood and are recognized as a valuable cell type in tissue engineering and cell therapy. Furthermore, hEnSCs, which have CD90 (a mesenchymal marker), CD105 (endoglin), CD44, CD146 (endometrial stem cell markers) and lack CD31 (Endothelial marker), CD34 (hematopoietic marker) and CD133 on the cell surface, are a new source of mesenchymal stem/stromal cells. Additionally, these cells can be encapsulated into self-assembling peptide nanofibers as a 3D scaffold for applications in the treatment of neurodegenerative diseases. Here, we describe a protocol to isolate hEnSCs from endometrium or menstrual blood. [摘要]  人类子宫内膜干细胞/基质细胞(hEnSCs)是从子宫内膜或经血中分离出来的,在组织工程和细胞治疗中被公认为有价值的细胞类型。 此外,在细胞表面具有CD90(间充质标记),CD105(内皮糖蛋白),CD44,CD146(子宫内膜干细胞标记)和缺乏CD31(内皮标记),CD34(造血标记)和CD133的hEnSCs是新的 间充质干细胞/基质细胞的来源。 此外,这些细胞可以封装成自组装肽纳米纤维作为三维支架应用于治疗神经退行性疾病。 在这里,我们描述了从子宫内膜或经血中分离hEnSC的方案。

【背景】正在研究细胞替代治疗,作为治疗神经退行性疾病(如阿尔茨海默病,中风和脊髓损伤)的新策略(Tavakol等人,2014b; 2015; 2016a和2016c)。子宫内膜干细胞/基质细胞(hEnSCs)可以从经血(Azedi等人,2014和2017)和子宫内膜分离。这些细胞具有脂肪形成(Khanmohammadi等人,2014),成骨(Darzi等人,2012)和软骨形成(Kazemnejad等人, >,2012)分化潜力。此外,这些细胞可以与支架组合用作组织工程应用中智能细胞支架的成分(Tavakol等人,2014a; 2016b和2017)。值得注意的是,诸如外来体,微泡和其他组分的细胞区室被认为比使用全细胞的治疗更安全。 ...

Primary Olfactory Ensheathing Cell Culture from Human Olfactory Mucosa Specimen
Author:
Date:
2017-05-20
[Abstract]  The human olfactory mucosa is located in the middle and superior turbinates, and the septum of nasal cavity. Olfactory mucosa plays an important role in detection of odours and it is also the only nervous tissue that is exposed to the external environment. This property leads to easy access to the olfactory mucosa for achieving various researches. The lamina propria of olfactory mucosa consists of olfactory ensheathing cells (OECs) that cover the nerve fibers of olfactory. Here we describe a protocol for isolation of OECs from biopsy of human olfactory mucosa. [摘要]  人类嗅粘膜位于中鼻甲和鼻中隔,鼻腔隔膜。嗅粘膜在气味检测中起重要作用,也是暴露于外界环境的唯一神经组织。该属性可以方便地进入嗅粘膜进行各种研究。嗅粘膜固有层由覆盖嗅神经纤维的嗅鞘细胞(OEC)组成。在这里,我们描述了从人类嗅粘膜活检中分离OEC的方案。

背景 嗅鞘细胞(OECs)是神经胶质细胞,其表达与星形胶质细胞和施旺细胞类似的各种抗原,例如胶质纤维相关蛋白(GFAP),S100-β,p75低亲和力神经生长因子受体,波形蛋白,巢蛋白和神经肽Y (Singh等人,2013)。嗅鞘细胞释放在中枢神经系统的细胞生长和粘连中起作用的不同神经营养因子和粘附分子(Pastrana等人,2007)。此外,这些细胞在受损的中枢神经系统的再生中起重要作用,例如治疗脊髓损伤和神经变性疾病(Novikova等人,2011)。我们选择OEC作为我们研究中的研究材料,因为它们具有诸如高迁移能力,可及源,鼻嗅粘膜干细胞分化和非致瘤性行为等优势性质(Huang et al。 ,2008; Escada 等人,2009)。该方案描述了从人类嗅觉粘膜样品中分离OEC的一步一步的过程。

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