Culture of Endometrial Explants and Peri-implantation Conceptuses to Monitor Synthesis and Secretion 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.
Estimation of Cell Number by Hemocytometry Counting Protocol 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.
Growing Adherent Cells on Coverslips or Multiwell Slides 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.
Plating Cells for Microinjection 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.
Static Culture of Postimplantation Embryos 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.
Worthington Tissue Dissociation Guide 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...