Initiation of Cell Lines
A wide array of established techniques is used to prepare stable primary cell cultures from human and animal tissues and to derive serially sub-cultured cell lines from these cultures.
Established cell lines are routinely obtained from a variety of sources and passaged in culture.
3D Organoid Culture
The facility can establish and passage organoid cultures from a variety of human and mouse primary tissues and tumors, as well as support labs performing these cultures with advice and reagents.
Mouse Embryonic Stem (ES) Cell Culture
The facility provides technical support, cells, reagents and supplies for culturing mouse ES cells, and performs transfections and cloning to establish germline-transmissible modified mouse ES cells to create knockout or knock-in mice.
Hybridoma Fusions and Antibody Production
The facility can produce hybridoma fusions, clone antibody-producing hybridomas, generate high titer monoclonal antibody (mAb)-containing culture supernatants, and bank hybridomas.
Immortalized B cells can be generated from human peripheral blood samples by transformation with Epstein Barr Virus (EBV).
Mycoplasma infestations of mammalian cell cultures can be detected by two methods in the Facility: 1) transferring their medium to indicator cultures of Vero monkey kidney cells and detecting cytoplasmic mycoplasma DNA by fluorescence microscopy or 2) using the Lonza MycoAlert Mycoplasma Detection Kit.
Preparation of Sterile Growth Media and Reagents
The CCF is a cost-effective resource for the preparation of complex media as well as more common media. Media and cell culture additives are available in a walk-in cold room and freezers that can be accessed 24/7 by self-serve and billing is logged to individual labs by a bar code reader system upon checkout.
Performance Testing of Fetal Bovine Serum
About every two years, approximately 500 liters of fetal bovine serum are purchased from a reserved production lot after being identified as optimal by performance testing within the Facility on seven sensitive cell lines. By testing and purchasing an optimal fetal bovine serum (FBS) batch in bulk, FCCC investigators have access to a consistent, quality-assured supply of FBS at the most cost-effective price.
Frozen Storage of Viable Cells (cell banking)
Viable cell samples are stored under liquid nitrogen (-196°C) in cryofreezers with a total capacity of approximately 123,000 computer-cataloged samples.
Cell Validation Support
The facility supports Fox Chase researchers by providing a wide array of common stock cell lines that were obtained directly from approved sources and by validation testing through STR marker profiling by IDEXX BioResearch. Currently, the facility has over 150 genetically-authenticated human and mouse cell lines available for distribution to FCCC researchers.
Immortalization of Mouse Embryo Fibroblasts (MEFs)
The facility immortalizes mouse embryo fibroblasts (MEF) by transfection with a vector encoding the simian virus large T antigen (SV40 TAg) oncoprotein to promote transformation.
Irradiated MEFs and 3T3-J2 Cells
Irradiated MEFs and 3T3-J2 are available to serve as feeder layers for ES and primary cell lines, respectively.