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However, the gene manipulation of T cells is extremely difficult, ordinary DNA plasmid is difficult to transfer, even virus mediated gene transfection operation, there are many difficulties, mainly reflected in several aspects:
1. Due to: 1) the instability of the receptor on the T cell membrane; 2) non-activated T cells have no ability to proliferate; 3) there are mechanisms in T cells that hinder reverse transcription,conventional lentivirus and retrovirus have weak infection ability to T cells, which requires repeated infection many times, and the conditions for experimental operation are very demanding.
2.T cells cultured in vitro are very sensitive and fragile, and lentiviral infection has a great impact on the state of the cells.
3. T cells are suspended cells, which further increases the difficulty of effective virus infection.
The viruses developed by Hanbio for the primary T cells are: T cell-specific lentivirus and adenovirus (Ads) for suspension cells.
1. The divided cells which from the immune system and hemopoietic system and other sources can effective infected;
2. Viral infection with human or mouse cell specificity;
3. Be used to build stable cell lines.
Cell: Human primary CD4+ T cell
Stimulation method: CD3/CD28 antibody and IL-2 activation of 48h
Virus: hTLv-GFP Lentivirus 1*108 TU/ml
Usage：MOI=50
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