GENE TRANSFER and SOMATIC CELL ENGINEERING

Sloan-Kettering Inst Can Research, New York NY

Investigators

Linked publications, trials & patents

Abstract

The Gene Transfer and Somatic Cell Engineering Core (GTS) supports the preclinical translation and clinical implementation of gene transfer studies at MSKCC. The supported projects are highly dependent on achieving efficient gene transfer in primary cells, including hematopoietic progenitor cells, T lymphocytes and dendritic cells. In the upcoming grant cycle, the GTS will mainly focus on the development and optimization of 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 patient cells in semi-closed systems in collaboration with the investigators for clinical trials utilizing genetically modified 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-closed systems; 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 in patient cells; 7. Cell banking, storage of viral stocks, plasmid DMA vaccine and clinical specimens. In addition, the GTS provides essential advisory and training functions for the generation of research grade reagents at the Center. Investigators are thus advised or trained on 1. How to optimize the transduction of various cell types; 2. How to construct recombinant gamma-retroviral and lentiviral vectors, plasmid DMA vectors, and shRNA encoding retroviral vectors; 3. What packaging cell lines to use; 4. How to transfect vector DMA in packaging cells and select producer cell lines; 5. What tests to perform to analyze gene expression; 6. How to titrate cell-free retroviral stocks by flow cytometry, Southern blot or real time PCR analysis. The GTS is thus a repository for numerous reagents and protocols that are made available to investigators at MSKCC. The centralization of cell transduction, vector production and plasmid DMA manufacturing in the GTS decreases the cost of clinical development, ensures high quality and consistency of molecular and cellular processes, and their availability to all investigators at the Center.