Mammalian Cell Culture Protocols

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. - [Read Adapting Cells to a Serum-Free Environment 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. - [Read Basic Techniques for Mammalian Cell Tissue Culture Protocol]

Culture conditions for various stable cell lines. - [Read Culture Conditions for Various Stable Cell Lines]

Protocol for ES / MEF cell culture and electroporation of targeting construct. - [Read ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol]

Covers general things you need to know to get started in cell culture work. Includes: Sterile Technique; Growth of Your Cell Culture; - [Read General Cell Culture]

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. - [Read Generation of Growth Curve 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. - [Read Germ Layer Explant Recombination Culture Protocol]

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. - [Read Growth of Primary Cell Culture and Viral Vector Handling Protocols]

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. - [Read Human Oesophageal 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. - [Read Human Thyroid Culture Protocol]

Information on Immortalized Mammary Epithelial Cells (IMEC). Includes: Background Information on IMEC; Culturing; Trypsinization. - [Read IMEC Maintenance]

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. - [Read Isolating Germ Cells from the Genital Ridge Protocol]

This procedure describes the isolation and culture of adult mouse cardiac myocytes from one heart. The isolation routinely yields approximately 1 million rodshaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from One Heart 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. - [Read Isolation of Adult Mouse Cardiac Myocytes from Two or More Hearts Protocol]

This protocol describes the isolation of fibroblasts from mouse, rat, and hamster embryos. - [Read Isolation of Fibroblasts from Mammalian Embryos Protocol]

Methodological details about sorting and purification of peripheral blood and decidual human NK subsets. - [Read Isolation of Human NK Subsets 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. - [Read Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol]

Protocol for osteoblast harvest (murine). - [Read Osteoblast Harvest (murine) Protocol]

Protocol describes trophoblast cell preparation. - [Read Preparation and Culture of Cytotrophoblasts Protocol]

Protocol for the preparation of NeoR murine embryonic fibroblasts. - [Read Preparation of NeoR Murine Embryonic Fibroblasts Protocol]

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. - [Read Protocol for Isolation, Cryopreservation, and Thawing of PBMCs]

Protocol for primary cultures of HUVECs. - [Read Protocol for Primary Cultures of HUVECs]

This protocol is optimized for erythroid cells, other cell types may require modifications. - [Read Protocol for Stable Lines]

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; - [Read Quality Control Considerations for Cell Culture]

Protocol to be used as reference to help standardization of both the isolation procedure and the quality of the cultures allowing a better comparison of the results obtained from different laboratories. - [Read Rat Chromaffin Cells Primary Cultures: Standardization and Quality Assessment for Single-Cell Assay]