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Tissue Culture Shared Resource at CU Cancer Center
 

Protein Production/MoAB/Tissue Culture


The Protein Production, Monoclonal Antibody, Tissue Culture Shared Resource (PPSR) supports basic and translational research at CU Cancer Center Members with a variety of services. The Shared Resource has over 200 authenticated human cell lines that are available for purchase at rates dramatically below those of commercial vendors. We can test your cell lines for mycoplasma, as well as offer authentication services on campus.

The Shared Resources has over 25 years' experience in the preparation of custom hybridomas using investigator-provided antigens (over one thousand hybridomas generated), large-scale growth of hybridomas, and purification of monoclonal antibodies (50 mg) have been producd for preclinical studies. 

We can grow small and large-scale (up to ten liters) preparations of cells for use in research, including for injection into animals. Investigators can also use either insect cells or mammalian cells as platforms for protein production. In support of insect cells, the shared resource can isolate and titer custom-made baculovirus expressing recombinant proteins of interest to researchers. We have also conducted large-scale transfection and cultivation of mammalian cells in support of large-scale preparation of recombinant proteins and monoclonal antibodies. 

The Shared Resource also has two Essen BioScience ZOOM IncuCytes for real time imaging of cell in culture. This platform can accommodate a wide variety of tissue culture vessels, although most investigators use 96 well plates for biological assays, and up to six different investigators can be accommodated in each instrument. The IncuCyte can quantitate cell proliferation using either phase contrast imaging or can directly quantitate cell number when cells expressing a nuclear GFP or RFP marker are used. Real time quantitation of apoptotic cells can be accomplished in living cultures without loss of cells using kits available in the Shared Resources. Quantitation of cell migration following wounding can be conducted in real time and quantitated using available image analysis software. Angiogenesis and neurite outgrowth can also be quantitated using this platform. 

Finally, specially designed 96 well plates can be used to measure chemotaxis of cells in real time. The scanning interval for image acquisition can be varied to meet the needs of the experimental design from as short as 20 minutes to once a day, although generally scanning intervals of 2 to 4 hours are used. Reagents for specialized assays are available on site. The Shared Resource sells a panel of cell lines with nuclear GFP or RFP markers to University investigators. In addition, they can assist in the preparation of new cell lines expressing nuclear GFP or RPF on a charge back basis. Finally, the Shared Resource offers an on-site service center stocked with culture media and related materials, as well as serum products, including large lots of tested fetal calf serum available at reduced price relate to existing market rates. Although many serum products are available, the Shared Resource purchased one large lot (700-1,000 bottles) of fetal calf serum that is subjected to rigorous on-site testing,; this lot is offered as a significant discount to commercial prices for comparable lots of serum.