Study overview: Automated spinoculation for gene transfer by lentiviral transduction. There is a growing need to close and automate manufacturing of chimeric antigen receptor (CAR) T cell therapies. CAR T workflows depend on ex vivo gene transfer by lentiviral transduction for

2023年7月31日 · For gene transduction into T- or NK cells by spinoculation, the cells were centrifuged with lentiviral or retroviral supernatant containing 8 µg/ml polybrene for 90 min at 1000–2000g at room ...

This protocol describes the use of MISSION ® TRC shRNA Lentiviral Particles particularly for long term silencing and phenotypic observation in suspension cells. The following protocol has been developed from the literature 1 for use in a 6-well plate with MISSION ® lentiviral particles.

2021年11月24日 · The effectiveness of lentiviral- and retroviral-mediated gene transfer has been shown to be significantly enhanced when a method called spinoculation is used [14,15,16,17,18,19,20,21,22]. During the process of spinoculation, both the viral vector and the cells are centrifuged together to aid viral binding to the target cells.

Lentiviral transduction is an effective method for creating a stable cell line with a DNA cassette of interest integrated into its genomic DNA, e.g. an sgRNA or gene expression cassette.

When transducing cells with lentiviral particles, a number of methods may be tried to enhance transduction efficacy such as spinoculation or prolonged incubation. High titer PGK-eGFP lentivirus was used to enable visualization of transduction efficacy by fluorescent microscopy.

This protocol is for transduction of suspension cells (Jurkat T cells, PBMC, PBL, B cells etc.) with lentivirus. To obtain the desired infection rate (low and high MOI; multiplicity of infection) in your target cells, serial

2014年2月27日 · We now describe a flexible and efficient approach involving virus-like particles containing simian immunodeficiency virus lentiviral gene product protein X and pseudotyping with CXCR4-tropic HIV...

2021年11月24日 · Closed system-bag spinoculation resulted in more efficient lymphocyte gene transfer than standard bag transductions without spinoculation. This method is effective for both retroviral and lentiviral vector gene transfer in lymphocytes and may be a feasible approach for gene transfer into other cell …

A combination of spinoculation and poloxamer-based chemical adjuvant increases the transduction of primary T-cells by greater than twofold. The combination of poloxamer-based and scFv-retargeted LVs increased transduction of CD30+ lymphoma cells more than tenfold, and has the potential to improve clinical protocols.