Adapting Cells to a Serum-Free Environment Protocol There are many ways to adapt cell lines to serum-free media. Five methods are presented that are designed for adapting hybridomas to a protein-free medium. These protocols may require some modifications for your particular cell line and conditions.
Basic Techniques for Mammalian Cell Tissue Culture Protocol Cultured mammalian cells are used extensively in cell biology studies; it requires a number of special skills in order to be able to preserve the structure, function, behavior and biology of the cells. This unit describes the basic skills required to maintain and preserve cell cultures: aseptic technique, medium characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells.
General Cell Culture Covers general things you need to know to get started in cell culture work. Includes: Sterile Technique; Growth of Your Cell Culture;
Generation of Growth Curve Protocol Generation of a growth curve can be useful in evaluating the growth characteristics of a cell line.
From a growth curve, the lag time, population doubling time, and saturation density can be
determined.
Germ Layer Explant Recombination Culture Protocol This protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos.
Growth of Primary Cell Culture and Viral Vector Handling Protocols The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents.
Human Oesophageal Culture Protocol This procedure describes a method for establishing short-term explant cultures of oesophageal mucosa. Adverse effects produced by exposure to radiation or test compounds can be detected as an inhibition of cell outgrowth.
Human Thyroid Culture Protocol This method enables the culturing of thyroid cells without loss of differentiation and medium change. It is potentially useful for the long-term study of drug effects on the thyroid gland.
IMEC Maintenance Information on Immortalized Mammary Epithelial Cells (IMEC). Includes: Background Information on IMEC; Culturing; Trypsinization.
Isolating Germ Cells from the Genital Ridge Protocol This protocol describes isolation of germ cells from the genital ridge of fetal mice from 11.5 days post coitum (dpc) onward. The germ cells can then be used for analysis, culture, or transplantation.
Isolation of Adult Mouse Cardiac Myocytes from Two or More Hearts Protocol This procedure describes the isolation and culture of adult mouse cardiac myocytes from two or more hearts. Includes
modifications for the digestion of two or more hearts in the same procedure and subsequent pooling of myocytes derived from the multiple hearts. The isolation procedure is performed by one or more technicians and routinely yields approximately 1
million rod-shaped myocytes per heart.
Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol This protocol introduces a method for maintaining mammalian cell culture suitable for treatment with siRNA. Transfection of cultured cells with siRNAs (see Transfection of siRNA Duplexes into Mammalian Cells) and downstream analysis of the knockdown cells.
Protocol for Isolation, Cryopreservation, and Thawing of PBMCs Cryopreserved PBMCs are a common specimen source for studies of immunological responses to vaccines,
immunotherapies, etc. The health and viability of cells recovered post-cryopreservation are of course
critical to the success and accuracy of immunological assays performed on them. This protocol standardizes PBMC isolation and cryopreservation techniques, specifically for the
assessment of thawed cells by cytokine flow cytometry.
Protocol for Stable Lines This protocol is optimized for erythroid cells, other cell types may require modifications.
Quality Control Considerations for Cell Culture Quality control considerations for cell culture. Includes: Provenance and Integrity of Cell Lines; Avoidance of Microbial Contamination; Environmental Monitoring; What to do in the event of contamination;