Adult nephroblastoma HANB cell strain and culturing method and application thereof
A technology for Wilms tumor and a culture method, which is applied to the field of adult Wilms tumor cell line and its culture, can solve the problems of difficulty in researching the mechanism of drug sensitivity and other problems, and achieves the effects of rapid growth and simple culture.
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Embodiment 1
[0038] Adult nephroblastoma HANB cell line, the preservation number is CCTCC NO: C201133, and its culture method is as follows:
[0039] 1. Culture and passage
[0040] Centrifuge the postoperative ascites from a patient with poorly differentiated anaplastic Wilms tumor at 1000 rpm for 10 minutes, discard the supernatant, and wash the precipitate with 1640 culture medium to obtain primary cells. 1640 culture medium is prepared from 1640 medium Cheng, 1640 culture medium is a commodity sold on the market, produced by American Life Tech Company, and sold by Beijing Dongge Biotechnology Co., Ltd.; the preparation method of 1640 culture medium is: 10g 1640 medium, 5.0g HEPES, 3.7g NaHCO 31. Dissolve 3g of L-glutamine in 1L of deionized water, filter through a 0.22μm microporous filter, sterilize and subpackage, and refrigerate at 4°C. On the ultra-clean workbench, place the primary cells in a cell culture flask, suspend them in 1640 culture medium with 20% newborn calf serum, and...
Embodiment 2
[0048] Adult nephroblastoma HANB cell line, the preservation number is CCTCC NO: C201133, and its culture method is as follows:
[0049] In step 1 of the culture method of adult nephroblastoma HANB cell line culture method, the primary cells are placed in culture flasks on the ultra-clean workbench, and cultured with 1640 containing 5% newborn calf serum. Liquid suspension, at 37°C, CO 2 Culture in a cell incubator with a volume concentration of 5% and a humidity of 95%, and change the 1640 culture medium containing 5% newborn calf serum every 24 hours until the cells grow to the plateau stage and complete the first stage of cell growth. For the second subculture, the cells grown to the plateau stage were subcultured; in the subculture step, digested with 0.22% trypsin aqueous solution, discarded the trypsin aqueous solution, and used newborn calf serum containing 1% mass fraction The 1640 culture medium was blown on the bottle wall to suspend the cells, and the suspension wa...
Embodiment 3
[0052] Adult nephroblastoma HANB cell line, the preservation number is CCTCC NO: C201133, and its culture method is as follows:
[0053] In step 1 of the culture method of adult nephroblastoma HANB cell line culture method, the primary cells are placed in culture flasks on the ultra-clean workbench, and cultured with 1640 containing 20% newborn calf serum. Liquid suspension, at 37°C, CO 2 Culture in a cell incubator with a volume concentration of 5% and a humidity of 95%, and change the 1640 culture medium containing 15% newborn calf serum every 24 hours until the cells grow to the plateau stage and complete the first stage of cell growth. For the second subculture, the cells that have grown to the plateau stage are subcultured; in the subculture step, digest with a trypsin aqueous solution with a mass fraction of 0.28%, discard the trypsin aqueous solution, and use a newborn calf serum with a mass fraction of 5% The 1640 culture medium was blown on the bottle wall to suspe...
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