Cell Culture Protocols

Protocol for culture of BEND cells (Bovine Endometrial Cells). Includes: Culture medium, Thawing Cells, Trypsinazation/ Splitting and freezing. - [Read Culture of BEND Cells (Bovine Endometrial Cells) Protocol]

Protocol used to study secretion of proteins and prostaglandins by endometrium from the cow, ewe, mare, bitch and other species. The technique is also useful for culture of peri-implantation conceptuses and placental tissues for metabolic labelling studies and to obtain conceptus secretory proteins for biological studies.The medium used is called Pig MEM, which is a modified minimum essential medium supplemented with non-essential amino acids, vitamins, insulin and additional glucose. - [Read Culture of Endometrial Explants and Peri-implantation Conceptuses to Monitor Synthesis and Secretion]

Protocol describes a method for estimation of mammalian cell number in a defined volume of medium using a hemocytometer. Automated methods using cell-counting devices such as those produced by Coulter are desirable when large numbers of individual samples are to be counted. - [Read Estimation of Cell Number by Hemocytometry Counting Protocol]

For low-resolution work, cells to be used for staining can be grown directly on regular tissue-culture dishes. It is a convenient method that does not require much preparatory work. - [Read Growing Adherent Cells Directly on Tissue Dishes Protocol]

Glass is an excellent substrate for most tissue-culture-adapted cells and is compatible with all fixing and staining solutions. Glass coverslips in tissue-culture dishes or in 24-well multiwell plates are suitable carriers, as are multiwell slides. For high-resolution studies, choose glass coverslips of the highest available grade; #1 or #1.5 coverslips are the appropriate thickness. - [Read Growing Adherent Cells on Coverslips or Multiwell Slides Protocol]

Information on the methods used to cultivate large amounts of cells for the purpose of obtaining an endogenous or recombinant product. - [Read Large-Scale Cell Culture Protocol]

Protocol describes a method for plating cells for microinjection onto etched coverslips. The coverslips for microinjection must be marked so that microinjected cells can be identified at time points after injection. - [Read Plating Cells for Microinjection Protocol]

Protocol describes static culture of postimplantation embryos, an alternative to the roller method. The static method is best suited to 6.0 to 7.0 days post coitum (dpc) embryos followed for 24 hours (7.0 dpc embryos) to 48 hours (6.0 dpc embryos) of development. It allows repetitive real-time observation with minimal handling of the embryo. It is especially useful if single or small groups of embryos need to be distinguished from each other. - [Read Static Culture of Postimplantation Embryos Protocol]

Includes information on: Cell Isolation Theory ( Tissue Types: Epithelial Tissue, Connective Tissue); Dissociating Enzymes (Collagenase, Trypsin, Elastase, Hyaluronidase, Papain, Chymotrypsin, Deoxyribonuclease I, Neutral Protease (Dispase), Trypsin Inhibitor (Soybean)); Cell Isolation Techniques: Working With Enzymes, Basic Primary Cell Isolation, Equilibration with 95%O2:5%CO2, Trituration, Enzymatic Cell Harvesting, Cell Adhesion and Harvesting, Trypsin for Cell Harvesting, etc... - [Read Worthington Tissue Dissociation Guide]

Protocol on [3H]Thymidine labeling. Protocol includes: Explant Culture Medium; 100X (cold) Thymidine; Dektol Developer; Fixer - [Read [3H]Thymidine Labeling Protocol]