The Gene Transfer and Somatic Cell Engineering Core (GTS) supports the preclinical translation andclinical implementation of gene transfer studies at MSKCC. The supported projects are highly dependent onachieving efficient gene transfer in primary cells, including hematopoietic progenitor cells, T lymphocytes anddendritic cells. In the upcoming grant cycle, the GTS will mainly focus on the development and optimizationof clinical cell engineering processes and on the implementation of clinical trials.
The specific aims of the GTS are to carry out and/or coordinate: 1. Expansion and transduction of patientcells in semi-closed systems in collaboration with the investigators for clinical trials utilizing geneticallymodified cells; 2. Generation and characterization of high-titer producer cell clones, master cell banks (MCB)for clinical studies; 3. Production and titration of 5 to 15 liter batches of clinical viral stocks in semi-closedsystems; 4. Production and biosafety testing of clinical grade plasmid DMA vaccine for immunization; 5.Detection of replication-competent retrovirus and other biosafety testing in cultured packaging cell clones(MCB), viral stocks and clinical specimen 6. Detection of oncoretroviral vector integration sites by LM-PCR inpatient cells; 7. Cell banking, storage of viral stocks, plasmid DMA vaccine and clinical specimens. Inaddition, the GTS provides essential advisory and training functions for the generation of research gradereagents at the Center. Investigators are thus advised or trained on 1. How to optimize the transduction ofvarious cell types; 2. How to construct recombinant gamma-retroviral and lentiviral vectors, plasmid DMAvectors, and shRNA encoding retroviral vectors; 3. What packaging cell lines to use; 4. How to transfectvector DMA in packaging cells and select producer cell lines; 5. What tests to perform to analyze geneexpression; 6. How to titrate cell-free retroviral stocks by flow cytometry, Southern blot or real time PCRanalysis. The GTS is thus a repository for numerous reagents and protocols that are made available toinvestigators at MSKCC.The centralization of cell transduction, vector production and plasmid DMA manufacturing in the GTSdecreases the cost of clinical development, ensures high quality and consistency of molecular and cellularprocesses, and their availability to all investigators at the Center.
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