Guides: BB 3870: Cell Culture Models Laboratory: Protocols

Protocols

Common Mammalian Cell Culture Protocols

Soft Agar Colony Formation Assay
A protocol from the National Library of Medicine. Tthe soft agar colony formation assay is a well-established method for characterizing the ability of cells to proliferate without a solid surface in vitro and is considered to be one of the most stringent tests for malignant transformation in cells.
Cell Culture Basics Instructional Videos
A collection of basic cell culture techniques and methods for passaging, freezing, and thawing cultured cells created by Gibco and provided on the ThermoFisher Scientific website.
Surface Areas and Guide for Recommended Medium Volumes for Corning Cell Culture Vessels
This guide gives the recommended medium volumes, approximate growth surface areas, and average cell yields for Corning disposable cell culture vessels.
Typical Hemocytometer Cell Counting Protocol from Thermofisher
A hemocytometer is a specialized glass slide with grid marks to visually count cells under a microscope and determine average cell density in a sample.
Cellometer Auto M10, Cellometer Auto T4 & Cellometer Auto T4 Plus User Manual
This manual provides descriptions of each automatic cell counter (by image analysis) and procedures for use.
General Scratch Assay Protocol from Axion Biosystems
The general outline for a scratch assay in which a small pipette tip is scraped along the bottom of a densely populated cell culture.
Thermofisher AlamarBlue Assay Protocol and Product Information
Thermofisher's informational packet on the use and properties of the AlamarBlue cell staining agent.
Example Resazurin Cell Viability Assay from Creative Bioarray
Living cells maintain a reducing environment within their cytoplasm and mitochondria, in which resazurin (blue and non-fluorescent) is reduced by dehydrogenase enzymes to form the red fluorescent dye resorufin. The amount of resorufin can be monitored by measuring fluorescence or absorbance, which is proportional to the number of living cells in the sample.
Sodium Alginate Preparation from a 2015 Study
"Sodium alginate preparation" and "Microsphere synthesis" are two particularly helpful methods in this published paper.
Sodium Alignate and PDMS (dimethylpolysiloxane) preparation from a 2014 Study
Read the methods section of this paper to learn how this group used alginate and PDMS to create environments for mammalian and algal cells.
Cell Culture of Human Keratinocytes
The section "Cell Culture and 3D (Static) Culture" may provide guidance for cell culture of other human cell types.
Promega's Cell Proliferation Assay Manual
The CellTiter 96® AQueous One Solution Cell Proliferation Assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays.

Other Protocols

SpringerProtocols (1980-2024)
SpringerProtocols is the world's largest collection of biomedical and life sciences protocols with over 65,000 articles. Based on tried and tested resources including Methods in Molecular Biology, SpringerProtocols includes provides current and alternative versions of protocols important for use in labs that don’t have the latest equipment. SpringerProtocols offers step-by-step laboratory instructions, lists of the necessary equipment and ingredients, and notes on troubleshooting and safety precautions. SpringerProtocols cover numerous subject areas including: • Biochemistry • Bioinformatics • Biotechnology • Cancer Research • Cell Biology • Genetics /Genomics • Imaging / Radiology • Immunology • Infectious Diseases • Microbiology • Molecular Biology • Neuroscience • Pharmacology / Toxicology • Plant Sciences • Protein Science (Coverage 1980 - 2024)

Schematic illustration of direct co-culture model verses the transwell (indirect) co-culture model approach.

Image source: Figure 3 from Hadi, N. S. A., & Stopper, H. (2025). Micronuclei as genotoxicity endpoint applied in the co-culture of two mammalian cell lines. Mutation Research - Genetic Toxicology and Environmental Mutagenesis, 901, 503839. https://doi.org/10.1016/j.mrgentox.2024.503839

CC BY 4.0