细胞技术

很多实验者都不bother自己洗手，酒精一喷就去实验了。我告诉大家，这是极端错误。个人经验是，你仔细洗手比喷酒精效果还好。最好的是肥皂/香皂仔仔细细的洗手，一直到手腕，指甲缝都要洗。洗两遍是一个不错的选择。然后手上喷酒精。有人穿半截袖的白大褂去做实验，每次我看到都想骂。白大褂袖口要长，最好是袖口扎紧的那种，如果不能的话，把袖口窝向手臂。裸露的手腕部分一定也要喷酒精。戴上手套后手请避免接触工作台外面，如果不得已接触，那 ...

在细胞和组织培养领域，从上世纪70年代起二维（2D）培养科学家已经看到其局限性，并且更多地关注三维（3D）培养的优点，目前越来越多的研究从细胞培养的平面环境中转变到三维培养。当前，虽然对基于细胞的效应研究和毒性测试中，制药工业如今最常用的依旧是2D方式，3D培养技术已在学术研究中被广泛应用。细胞增殖，分化和代谢等生理活动都严重受到微环境的影响。当前细胞生物学研究大多还是在二维平面培养进行，这种平面培养、生长方式与 ...

调节性T细胞(Regulatory cell ,简称Treg )是一类控制体内自身免疫反应性的T细胞亚群。目前有越来越多的学者投身于Treg的研究当中，但对于Treg细胞的获取，您真的知道很多吗？科学家们研究发现当细胞受到损伤后，会释放NAD+（Nicotinamide Adenine Dinucleotide），而游离的NAD+能够与细胞上表达的酶ARTC2结合；ARTC2会将ADP-核糖转移到细胞表面蛋白而激活该蛋白，比如P2 ...

养了很久的细胞，有一些经验和体会总结一下，和大家分享一下。关于如何把细胞养的形态很好更漂亮。

1.不同的细胞，喜欢的环境是不一样的。

这个不仅仅是说培养基的不同，还有就是细胞生长的空间密度问题。

有一些细胞是数量多一点比较好生长，生长状态也比较好。这种一般是属于生长速度慢的细胞。譬如内皮细胞。而有一些是细胞是数量少一点细胞状态会生长的比较好，譬如巨噬细胞和某些肿瘤细胞。尤其是巨噬细胞，生长速度非常得快，贴壁速度很快，所以传代时 ...

1，天然培养基血清、组织提取液等血清：主要是小牛血清、胎牛血清营养成分--白蛋白、纤连蛋白、球蛋白、转铁蛋白、蛋白酶抑制剂、生长因子等蛋白质和多肽；40-80mg/ml氨基酸；0.01-1.0vmol/L 尿素； 170-300ug/ml胆固醇、脂肪酸、磷脂等类脂；2-10mg/ml葡萄糖、乳酸、丙酮酸等碳水化合物；1-2mg/ml钙、氯、铁、钾、钠、锌等无机物；014-0.16mol/L胰岛素、甲状腺素等激素；0.1-200nmol/L维生素A、叶 ...

Cell-Adhesion AssaysDennis F. Kucik and Chuanyue WuSummaryOne of the most important properties of cells that are derived from multicellularorganisms is their ability to adhere to extracellular matrix proteins or other cells. Analysisof cell–extracellular matrix and/or cell ...

PART II：BASIC CELL MIGRATION AND RELATED ASSAYSvol.294 Guan J.-L. (ed.) Cell Migration-Developmental Methods and Protocols Chapter 3Analysis of Cell Migration Using the Dunn Chemotaxis Chamber and Time-Lapse MicroscopyClaire M. Wells and Anne J. RidleySummaryThe directed mig ...

PART II：BASIC CELL MIGRATION AND RELATED ASSAYSvol.294 Guan J.-L. (ed.) Cell Migration-Developmental Methods and Protocols Chapter 3Wound-Healing AssayLuis G. Rodriguez, Xiaoyang Wu, and Jun-Lin GuanSummaryThe wound-healing assay is simple, inexpensive, and one of the earliest ...

PART II：BASIC CELL MIGRATION AND RELATED ASSAYSBoyden Chamber Assayvol.294 Guan J.-L. (ed.) Cell Migration-Developmental Methods and Protocols Chapter 2SummaryThe Boyden chamber assay, originally introduced by Boyden for the analysis ofleukocyte chemotaxis, is based on a cha ...

Cell Migration——An Overviewvol.294 Guan J.-L. (ed.) Cell Migration-Developmental Methods and Protocols Chapter 1SummaryCell migration is an essential process for normal development and homeostasis thatcan also contribute to important pathologies. Not surprisingly, th ...

Native Promoter Reporters Validate Transcriptional TargetsMethods in Molecular Biology v468. chapter 9Otto Schmalhofer, Simone Spaderna ,and Thomas BrabletzAbstractThe transcriptional activator b-catenin is the key mediator of the canonical Wnt signaling pathway ...

Assaying b-Catenin/TCF Transcription with b-Catenin/TCFTranscription-Based Reporter ConstructsMethods in Molecular Biology v468. chapter 8AbstractTranscription-based reporters have been instrumental in characterizing the Wnt/b-catenin signalingpa ...

Detection of b-Catenin Localization by ImmunohistochemistryMethods in Molecular Biology v468. chapter 7Abstractβ-catenin is a widely expressed 90-kDa protein with dual functions in cell adhesion and Wnt signalling. At the membrane, β-catenin forms complexes with E-cadherin ...

Detection of Cytoplasmic and Nuclear Localization of Adenomatous Polyposis Coli (APC) Protein in CellsMethods in Molecular Biology v468. chapter 6AbstractThe adenomatous polyposis coli (APC) tumour suppressor gene is mutated in the majority of colon cancers.APC is a multi-dom ...

Inhibition of Glycogen Synthase Kinase-3Methods in Molecular Biology v468. chapter 5AbstractThere are two homologous forms of glycogen synthase kinase (GSK)-3, GSK-3a and GSK-3b, whichplay overlapping roles in the regulation of Wnt, Hedgehog, and insulin pathways, as well as the act ...

AbstractGlycogen synthase kinase (GSK)-3 is a key signalling intermediate in the action of Wnts. This protein kinaseis ubiquitously expressed and has high inherent activity but is inhibited by activation of Wnt signalling oractivation of growth factor receptor tyrosine kinases (e ...

AbstractRecombinant expression of secreted Frizzled-related proteins (sFRPs) in mammalian expression systemsis a convenient source of these proteins for biological studies. Yields of protein vary; screening of clonallines for high expression is usually worthwhile. Hep ...

Wnt proteins and their signaling cascades are involved in a wide variety of developmental processes, andderegulation of this pathway is frequently associated with tumorigenesis. Unlike many other growthfactors, Wnts long eluded biochemical purification, in large part because of their hydrophobic nature,which is imparted by one or more lipid modifications (1 – 3) . Here I describe a complete protocol thatoutlines the purification process for Wnt proteins. While this protocol has not been applied

Embryonic development of multicellular organisms is an incredibly complex process that relies heavily on evolutionarily conserved signalling pathways to provide crucial cell–cell communication. Typically, secreted signalling proteins such as Wnts, BMPs, and Hedgehogs released by one cell population will trigger concentration-dependent responses in other cells located some distance away. In adults, the same signalling pathways orchestrate tissue renewal in organs such as the intestine and ski

树突状细胞培养操作步骤准备补充剂将补充剂混合物在 15~25℃ 解冻。在无菌条件下用移液枪小心地将补充剂混合物混匀。然后，把补充剂混合物全部加到基础培养基中。盖上瓶子，轻轻混匀直到形成均一的混合物。DC生成培养基所附带的相应的细胞因子组合包含有成分A和B，它们都是以100X 的原液状态运输的。注意：此时不要将化合物B加入培养基中。DC基础培养基不带细胞因子，因此使用时必须另外自行添加。新鲜分离的单核 DC 细胞的传代对于新鲜分离自 ...