细胞技术

记者从中科院广州生物医药与健康研究院获悉，由该院博士赖良学领衔的研究团队，日前将人类成纤维细胞直接转分化成为神经元限制性前体细胞。分化后的细胞能在体外培养条件下大量增殖，并且仅特异分化为神经元，而不会产生胶质细胞。这项技术将可能为采用细胞替换疗法治疗人类神经退行性疾病提供一个新的细胞来源。相关研究成果近日在线发表于《生物化学》杂志。据介绍，当前许多研究用成纤维细胞生成神经干细胞。神经干细胞可分化为神经元和胶质 ...

1月，Haruko Obokata在描述自己的新研究成果。图片来源：Kyodo1月至今，全世界科学家都无法复制震惊世人的干细胞研究成果，该研究称，简单加压就能将成体细胞回复成新细胞，这些细胞与在早期胚胎中发现的干细胞类似。现在，《科学》杂志获悉，在相关论文发表之前，参与撰写这两篇文章的一些实验室并未尝试重复该技术，其中只有两个实验室表示他们能够产生所谓的STAP细胞。这场纷争出现自两个月前，当时干细胞专家Haruko O ...

一、实验背景丙型肝炎病毒（HCV）是引起丙型肝炎的罪魁祸首，全球有 1.3 亿人慢性感染HCV并逐步 发展成肝硬化和肝癌，但就其致病机理却并不像其他肝炎病毒（如 HBV）一样清晰，在HCV进入肝细胞的过程中，一系列的受体识别和相互作用的参与其中，还包括由网格蛋白(clathrin)参与的内吞作用，但目前除 了受体之外，感染性 HCV 入侵的所需的其他的细胞分子还知之甚少。二、实验方法通过筛选了包含靶向标记 140 个细胞膜流通蛋白 siRNA 的文库， 以期找到 H ...

一、酶消化法1、胰酶。这是用得最多的。一般浓度在0.25-0.5%。作用时间根据细胞种类、作用温度等因素而变化很大，从几分钟到几十分钟不等。0.25%的胰酶作用于单层贴壁的细胞，在37度条件下，一般消化1-5分钟就足够了。终止是用血清。主要作用于细胞间。配制时不能用含钙、镁的平衡液，否则影响活性。保存于-20度。2、胶原酶。这种方法比较少，一般是用原代培养时，从组织消化下细胞。这种方法作用温和，对细胞损伤较小，但是，价格也较贵。中 ...

作为一只刚进实验室的菜鸟把自己遇到的东东总结一点发出来，抛砖引玉吧，为即将进实验室的朋友些资料，同时欢迎老鸟们指正。1、在实验室登记领试剂和药品pbs 1640 Gbico的FBS 台盼蓝 计数板 酒精灯 胶塞 封口胶 消毒的喷剂 记号笔 镊子 标签 饭盒 消毒时包饭盒的布 冻存管 冻存剂 离心管 培养瓶 6孔板 96空板 吸管 吸球。pbs 1640 Gbico的FBS 我都是买的现成的，就是那些实验室老师用原料配好了来卖的那种，我是害怕自己本来就是一个新手，为省那钱自己配，别污染了多的事情都来了， ...

自噬是近年来很热门的领域，做一下系统的介绍或讨论，内容：1） 什么是自噬？包括自噬的定义、形态学特征、分子基础、调控等2）自噬的意义自噬与细胞存活、细胞死亡、疾病、衰老等的关系3）怎么研究自噬？主要谈谈怎么证明细胞发生了自噬，即自噬的判断标准和各种研究自噬的方法及局限性。一、自噬的过程——从一张图片开始步骤1：细胞接受自噬诱导信号后，在胞浆的某处形成一个小的类似“脂质体”样的膜结构，然后不断扩张，但它并不呈球形，而是扁平的，就 ...

本视频主要介绍了用慢病毒腺病毒进行细胞转染之实验前期准备工作。用慢病毒腺病毒进行细胞转染之实验前期准备工作（真人示范）

一、方法根据1999年lutz的方法，做了一些改进，培养DC很多人用的是高浓度的细胞，高浓度的因子，但是这样产量不高，我每一次分离一只小鼠的骨髓，大约可以得到107个细胞，分成10盘，每盘10 ml培养基，用低浓度的细胞因子，这样可以大大提高产量，每次能培养得到undefined107个DC左右，流式细胞术检测：细胞纯度应该大于90%，这个方法还是很好用的。二、具体步骤1. C57Bl/6小鼠2. 处死小鼠，75%酒精浸泡10 min。3. 解剖取出小鼠股 ...

细胞培养（特别是传代细胞）被支原体污染是个世界性问题，在细胞培养中支原体感染发生率达到63%,只要有一个具有增殖能力的支原体就能引起培养体系的污染,支原体则像慢性毒药一般，杀细胞于无形中，据统计，超过25%的细胞系中感染了支原体，研究者又不能通过可视法对其进行检测，而且它对细胞的生长率影响较小，也不易引起注意，所以研究者并不知晓。支原体污染发生后危害巨大，能改变细胞的DNA,RNA及蛋白表达，改变细胞、篡改数据，造成 ...

The liver is composed of hepatocytes, endothelial, Kupffer, fat-storing, and pit cells. This chapter describes a technique for isolating hepatocytes, which represent 60–65% of the cells of the liver and about 80% of the volume of the organ (since they are larger than the other cells) (1–3). Two years aft ...

Macrophages throughout the body function to phagocytose pathogens, tissue debris, or foreign particulates. For example, lung macrophages have frequent contact with inhaled pathogens or inert particles and have developed interesting and diverse responses to these challenge ...

Eukaryotic cells are electronegative when compared to the surrounding environment. This negative charge is called the transmembrane potential (TMP) and is partly caused by concentration gradients of K+, Na2+, and Cl− ions across the cell membrane. For purposes of cell osmolarity and pH bal ...

The estimation of the viability of a cell population by flow cytometry is based on a simple yet powerful principle: Dead cells leak. Cells that die via the necrosis pathway, in contrast with apoptosis, rapidly lose membrane integrity (1). All of the different methods for evaluating viability are ba ...

Progression of cells through successive phases and checkpoints of the cell cycle is maintained by sequential phosphorylation of different sets of nuclear and cytoplasmic proteins by cyclin-dependent kinases (CDKs) (1–12). By activating their partner CDKs and targeting them to the r ...

The application of flow cytometry for the study of cell surface antigens has evolved significantly in the last 20 yr. One of the factors involved is the continuing development of a vast array of monoclonal antibodies specific for cell-associated antigens. Another important factor is the dev ...

The increased utilization of flow cytometry for the study of cell-associated antigens has paralleled the continuing development of new monoclonal antibodies (MAbs) specific for cell-associated antigens. Availability of these reagents, combined with the introduction of new ...

The current methods commonly employed to detect cytokine production have several drawbacks. Bioassays are not necessarily cytokine-specific in that they measure functional properties. The production of supernatant cytokine protein can be readily measured by enzyme-linked ...

The mere presence of terminal deoxynucleotidyl transferase (TdT), a DNA polymerase, in leukemic cells provides no help in assigning these blast cells to a particular cell lineage (1). Differential levels of TdT gene transcription, however, result in diagnostically significant expr ...

The development of simultaneous multiparameter analysis protocols has facilitated the use of flow cytometry in many diverse and complex research and clinical programs. This effort has been advanced by the availability of a wide range of fluorochrome-conjugated antibody reagents ...

Many types of cancer and genetic diseases are characterized by chromosomal aberrations. Conventional cytogenetics, the analysis of banded metaphase chromosomes, can be very time consuming and, in many cases, marker chromosomes cannot be identified by their banding pattern alone. F ...