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The Bioreagent and Cell Culture Core Facility supports cancer related research that requires in vitro experiments, provides reagents for cell culture and prepares Bioreagents that are produced by cells grown in the facility. Recently, the Cell Culture Core facility was reorganized to introduce new services of producing Bioreagents that were identified as high priority by the investigators. We now grow suspension cultures of hybridoma cells in large quantities and produces purified monoclonal antibodies. A recently acquired New Brunswick BioReactor made the growing of cells in large quantities possible at substantial savings. We are in the process of developing and evaluating lipofectin preparations for cell transfection at highly reduced costs. Additional available services include the preparations of specialized serum products, LB medium, matrix coated culture dishes, and AmpicillinR plates. Studies will be conducted to evaluate the feasibility of producing recombinant proteins, cytokines and growth factors.
We continue to provide established services of cell culture media preparations in large batches and their distribution at six convenient locations, testing for mycoplasma contaminations, drug testing using cytotoxicity and cell proliferation assays, bulk purchase of tissue culture additives, growing attached cells in large quantities using roller bottles, and assisting in all techniques related to cell cultures.
Cell Culture Media
Cellgro formulations
For information regarding in-house media formulations, please contact Nily Harel or Antranik Shahinian.
Mycoplasma Test
Mycoplasmas are the smallest and simplest prokaryotes which have long been recognized as common contaminants during the continuous cell cultures. Mycoplasmas depend on their hosts for nutrients and their presence may go unnoticed for months. As the mycoplasma competes with the cells for nutrients, the infected cultures may have reduced rates of proliferation, change in cellular responses to the drug treatments, altered cell surface signal and transport mechanisms, gene expression, and modified physiological functions.
Up to now about 118 species of mycoplasma have been reported, of these species about 10 are found as the most common culture contaminants. We have validated our detection assays using 15 uninfected cell lines as negative controls and 5 contaminated cell lines using three different methods: fluorescent staining, PCR, and enzymatic assays.
The assay used in our facility is based on the detection of a unique enzyme that is found in broad varieties of mycoplasma species (25 of 25 species tested). The assay measures energy generated by the enzyme from a specific substrate which consequently activates the enzyme luciferase. Luciferase activity is detected by luminescence emission from the oxidation of luciferin. This assay is very sensitive and highly specific for mycoplasma (virus, yeast, bacteria, and other lower microorganisms do not have it) and virtually all mycoplasma species express this enzyme. The sensitivity approaches that of PCR assay. If necessary, the Core Facility however will also perform the PCR based assay, but this assay may not detect all common species of mycoplasma.
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Contact us for further information.
Contact us for further information.
USC/Norris Comprehensive Cancer Center
1303 Mission Road, CRL-216
Los Angeles, CA 90033
Zoltan Tokes, PhD
Director
Telephone: (323)224-7775
Email: ztokes@usc.edu
Nily Harel
Research Lab Specialist
Telephone: (323) 224-7792
Email: harel@usc.edu
CRL-216
Telephone: (323) 224-7792
Norris 5323
Telephone: (323) 865-0527
E-mail: CoreCellCulture@ccnt.usc.edu