细胞技术

对骨髓基质干细胞进行改造使其在保持快速生长的同时向成骨细胞方向分化是目前国内外研究的热点。实验发现成熟的成骨细胞可以通过细胞间的直接接触作用促进骨髓基质干细胞增殖 此外成骨细胞可产生多种生长因子促进其骨髓基质干细胞的增殖、分化 。由于骨髓基质干细胞通过诱导向成骨细胞分化后难以再与原有的成骨细胞区分开因此本实验将骨髓基质干细胞与成骨细胞以1∶1 的比例直接混合培养同时将成骨细胞和骨髓基质干细胞在与混 ...

细胞的分裂指数一、原理体外培养细胞生长、分裂繁殖的能力，可用分裂指数来表示。它与生长曲线有一定的联系，如随着分裂指数的不断提高，细胞也就进入了指数生长期。分裂指数指细胞群体中分裂细胞所占的百分比，它是测定细胞周期的一个重要指标，也是不同实验研究选择细胞的重要依据。二、仪器、用品与试剂1、仪器与用品：CO2培养箱、普通显微镜、细胞培养血、盖玻片、吸管2、试剂：培养液、胰酶、甲醇、冰醋酸、Giemsa ...

Purpose and BackgroundsCHO lec 3.2.8.1 cellsCHO Lec 3.2.8.1 cells have four independent mutations in the N- and O- glycosylation pathways (Stanley 1989). N-linked carbohydrates produced by CHO Lec 3.2 ...

PurposeThis protocol describes how to isolate human platelets from whole blood. Isolated platelets are used for static adhesion assays for flow chamber assays flow cytometry measurements etc.Materials ...

地高辛（DIG）产品选择指南――选择合适的标记方法标记DNA探针如果您有足够量的高纯度模板，建议您采用随机引物标记方法的DIG High Prime DNA Labeling and Detection Starter Kit I 和II。这两个试剂盒包含了标记、封闭、杂交和检测所需要的所有试剂，是真正意义上的一站式服务的试剂盒。二者之间的差别在于检测方法的不同：Starter Kit I采用了显 ...

小提.pdfplasmid.pdfVigoFect.pdfmatrigel.pdf ...

MATERIALSMedia: Heat inactivated FBS (56°C x 30 min) PBS + 2% heat-inactivated FBS IMDM + 20% heat-inactivated FBS Viral transduction: Iscove's + 20% heat inactivated FBS plus 100 ng/mL each of SCF IL ...

Reservations:Schedule time on a cytometer (FACS or Calibur) by filling in the calendar at the flow lab. Don't grossly overbook since we have to pay for both reserved time and the time actually used (w ...

NoteThis protocol describes semi-automated cell counts using fluorescently labeled cells a hemocytometer and ImageJ software. The hemocytometer is not needed if you have already calibrated the areas ...

Materials 2x HEPES: 8 g NaCl 0.105g NaHPO4 6.5 g HEPES H2O to 500 mL pH 6.95 to 7.10 (try a range) 2M CaCl2: store in aliquots at -20°C DNA 4 mg/mL Procedure (The volumes can be scaled up as necessar ...

MaterialsWildtype MoMLV virus aliquot. Stored at -80ºC.Medium: DMEM + 10% FBSNIH 3T3 TK- cellsXC cellsPolybrene 1000x stock = 4 mg/mL sterile filtered.U.V. light source on a stand.ProcedureDay 0: T ...

1、将转染后的细胞按照一定的比例接种到6孔板中（一般选用1：10和1：100两个梯度），分别采用1200mg/ml，1000mg/ml和800mg/ml的G418进行筛选；以转染携带EGFP质粒载体的细胞作为阳性对照，以未转染质粒载体的细胞作为阴性对照；2、每3～4天换液一次；3、筛选20～30天，即可得到稳定表达细胞株；4、传代后，以一定的G418浓度维持培养； ...

The plasticity of differentiated adult cells could have a great therapeutic potential but at the same time it is characteristic of progression of serious pathological states such as cancer and fibrosi ...

1. 2 稳定转染细胞株的构建原理：稳定转染，就是转染的质粒DNA整合到宿主细胞染色体上，使宿主细胞可长期表达目的基因及蛋白。需要在瞬时转染基础上对靶细胞进行筛选或者应用高转染效率的病毒，根据不同基因载体中所含有的抗性标志选用相应的药物进行细胞传代，从而得到可稳定表达目的基因的细胞系。应用： 观察目的基因及其表达蛋白在某种细胞中的功能（稳定，可长期进行研究）一般流程：1） 筛选浓度测定： ...

OverviewCount the number of viable cells in a culture via dilution plating. The following assumes E. coli cells but it should apply to any type of cells. MaterialsLB agar plate LB media for dilution 8 ...

Cell cycle analysis of Escherichia coli cells C period = the time for a round of chromosome replication D period = the time between the end of a round of chromosome replication and cell division Deter ...

OverviewThis protocol is a variant of the Hanahan protocol using CCMB80 buffer for DH10B TOP10 and MachI strains. It builds on Example 2 of the Bloom05 patent as well. This protocol has been tested o ...

MethodThaw TSS cells on ice. Add DNA pipette gently to mix (1μl of prepped plasmid is more than enough). Note: If you are adding small volumes (~1μl) be careful to mix the culture well. Diluting the p ...

CuratorsJingyang Chen Seattle Biomedical Research Institute 307 Westlake Ave N Suite 500 Seattle WA 98109 USA. jingyang.chen@sbri.org Anyone should feel free to add themselves as a curator for this co ...

IntroductionApoptosis is a normal physiological phenomenon put forward by Kerr . It plays an important role in embryonic development maintenance of tissue homeostasis and pathology and therefore has b ...