Closed type lentiviral vector culturing device and lentiviral vector culturing method

Abstract

The invention discloses a closed type lentiviral vector culturing device and a lentiviral vector culturing method. The culturing device consists of a box body, a valve, a connecting pipe, a Q-syte diaphragm closed type needle-free joint, a Luer-Lock injector, a filter and a closed bolt, wherein the box body is formed by closely jointing an upper box piece and a lower box piece; the upper box piece and the lower box piece are fixed under a rigid frame; the central parts of the upper box piece and the lower box piece are sunken towards the outer side of the box body, so that a rectangular containing cavity is formed; variable cross-section cylindrical channels are respectively arranged at the left side and the right side of the box body, and a mother slot connecting tube is fixed in each channel; a joint of the Luer-Lock injector is a spiral joint. Before the culturing device is used, culture mediums of transduction plasmids, 293 T cells and Huh 7.5.1 cells, primary hepatocyte culture mediums and primary hepatocyte tissue are firstly prepared; the culturing device is used for culturing so as to obtain lentivirus supernatant; primary hepatocytes are used for carrying out transfection on the lentivirus supernatant; the lentiviral vector is finally obtained after being cultured for a certain number of days.

Description

technical field [0001] The invention relates to a closed lentiviral vector culture device and a culture method. Background technique [0002] Transgenic expression of mammalian proteins may convert normal somatic cells into a tumorigenic state, for example inactivation of the p53 and Rb pathways leads to expression of dominant-negative p53 protein (p53DD), an activated cyclin-dependent kinase (CDK ) / cyclin complex (cyclinD1 / CDK4 R24C ) with activation of the proto-oncogene, Ras, and hTERT pathways through the oncogenic expression of the proto-oncogene (c-Myc T58A ), H-Ras (H-Ras G12V ) and hTERT, can drive transformation of different human cells into tumorigenic states. Studies have reported that normal cells can undergo cancer-causing transformation using the combination of oncogenes described above, suggesting that normal liver cells may be genetically engineered to help discover the normal transformation process in human cancer when it returns to host liver cells Tumo...

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Problems solved by technology

However, the culture devices used in current biomedical experiments still cannot protect the experimenters well

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