This page introduces tissue culture methods for in vitro growth of cells. Cell culture techniques can vary enormously both technically and in quantities (range from less than 1 ml to sevreal 1000 ml). In general,there are 2 types of cell culture: primary (derived directly from a living organism) or cell lines (stable established cells which proliferate in culture). Both types of culture have pros/cons and employ similar practical sterile techniques.
For cell lines the links below will take you to a page with a full description and images of individual cell lines. Many of the lines (if not generated and a gift from specific Labs) are available from either ATCC or ECACC, the 2 large cell line repositories in USA and Europe respecively. The Lab also has a number of mouse hybridoma cell lines producing specific antibodies.
If tissue culture is a new topic to you, begin by looking at ANAT3231 Lab 2- Introduction to Tissue Culture
Page Links: Introduction | Primary Culture Overview | Cell Line Culture Overview | Laboratory Facilities | Lab Tools | Cell Sources | References | Commercial Resources
Primary cell cultures are derived from rat, mouse and human in the Lab. The majority of cultures focus on the growth of either neurons and/or glia. Cultures can be generated from embryonic or adult: cortex, retina, spinal cord, dorsal root ganglia, sympathetic ganglia. Depending on the preparation technique either neurons, glia or neurons and glial cultures can be generated.
The advantages of primary cultures are that the cells have not been "modified" in any way (other than enzymatic or physical dissociation).
The disadvantages of primary cultures are the mixed nature of each preparation, limited lifespan of the culture and the potential contamination problems. In some cases these cells can also be stored frozen for future use. Remember that in the adult (except for olefactory and ventricular) neurons are post-mitotic and will not proliferate unless transformed.
All primary cell culture experiments using animals need Animal Ethics approval and must comply with Australian Code of Practice for the Care and Use of Animals for Scientific Purposes, National Health and Medical Research, Council (6th edn, 1997). (More? UNSW Animal Care and Ethics Committee)
All primary cell culture experiments using human material need Human Ethics approval. (More? National Statement on Ethical Conduct in Research Involving Humans | UNSW Human Ethics)
For cell lines the links below will take you to a page with a full description and images of individual cell lines. Many of the lines (if not generated and a gift from specific Labs) are available from either ATCC or ECCC, the 2 large cell line repositories in USA and Europe respectively. The Lab also has a number of mouse hybridoma cell lines producing specific antibodies.
3T3 | B35 | C2C12 | C17-2 | HT4 | IGR3 | NT2 | PC12 | RGC5 |
Download PDF file of CBL Cells (36 Pages, 570 Kb)
The laboratory has OGTR certified PC2 tissue culture facilities available for use. Long-term cell storage in liquid nitrogen and short-term -80C freezer.
Below are links to useful resources, if someone already has a good tool, why reinvent the wheel? (links don't work? ...internet links change)
Using a Counting Chamber from Rice University.
Phosphate (Sodium) buffer Chart from Grain Genes.
The two suppliers below are the best source for fully characterised and tested cells and cell lines.
ATCC (USA) ATCC "... global nonprofit bioresource center that provides biological products, technical services, and educational programs to private industry, government, and academic organizations around the world. Our mission is to acquire, authenticate, preserve, develop, and distribute biological materials, information, technology, intellectual property, and standards for the advancement, validation, and application of scientific knowledge."
Bacteria, Bacteriophages, Cell Lines and Hybridomas, Filamentous Fungi and Yeast Plant Seeds, Protozoa and Algae, Viruses and Antisera.
Postal: ATCC, P.O.Box 1549, Manassas, VA 20108 USA Phone: (703) 365-2700 E-mail: news@atcc.org
ECACC (Europe) ECACC "The European Collection of Cell Cultures (ECACC) was established in 1984 as a cell culture collection to service the research community and provide an International Depository Authority recognised patent depository for Europe." "The collections currently hold over 40,000 cell lines representing 45 different species, 50 tissue types, 300 HLA types, 450 monoclonal antibodies and at least 800 genetic disorders."
Phone: +44 (0)1980 611315 Fax Number: 44 (0)1980 612512 Email Address: ecacc@hpa.org.uk
Mather JP. Making informed choices: medium, serum, and serum-free medium. How to choose the appropriate medium and culture system for the model you wish to create. Methods Cell Biol. 1998;57:19-30. Review.
Li Y, Powell S, Brunette E, Lebkowski J, Mandalam R. Expansion of human embryonic stem cells in defined serum-free medium devoid of animal-derived products. Biotechnol Bioeng. 2005 Sep 20;91(6):688-98.
Butler M. Animal cell cultures: recent achievements and perspectives in the production of biopharmaceuticals. Appl Microbiol Biotechnol. 2005 Aug;68(3):283-91.
These commercial links are for educational purposes only and do not reflect endorsement of a specific product.
BD Biosciences Websitehttp://www.bdbiosciences.com/discovery_labware
Promega Websitehttp://www.promega.com/
Serotec Websitehttp://www.Serotec.com/
There are many different uses and methods for growing cells in culture. This page is an introduction to the methods used in our lab.
Please email Dr Mark Hill if you wish to make a comment about this current project.