Product Series
Immunity
Primary neuroblastoma Conditional reprogramming medium is a medium developed to promote the growth of primary human neuroblastoma cells in vitro. It is a sterile liquid mixing system containing essential and non-essential amino acids, vitamins, organic and inorganic compounds, hormones, growth factors, trace minerals, and more. The medium product is based on a bicarbonate buffer system and has a pH of 7.4 when balanced in a 5% CO2, 37 ° C incubator. The formulation of this medium can provide a suitable culture environment for human neuroblastoma primary cells in vitro and selectively promote the growth of human neuroblastoma primary cells in vitro. The purpose of short in vitro culture cycle, controllable cost and convenient operation was realized, and the success rate of primary cell culture of tumor tissue was improved.
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Product Introduction
1. Neuroblastoma primary cell culture
The obtained primary cells of human neuroblastoma were inoculated into appropriate culture vessels according to the cell number in the instructions, and NIH-3T3 cells irradiated by gamma rays were added (gamma-NIH-3T3 was used as trophoblast cells, and the number of trophoblast cells was added in the instructions). The primary cells were not moved for 2 to 3 days after the initial inoculation (which is conducive to cell adhesion). During the culture process, if the medium color turns yellow but the cells are not overgrown, the solution can be half-changed. When the cells are not overgrown but the gamma-NIH-3T3 cells are insufficient, the gamma-NIH-3T3 cells should be properly supplemented (generally half of the initial number).
2. Neuroblastoma primary cell passage
The clones and neuron-like cell proliferation were observed under microscope, and the confluent degree was 85-95%. The cells were removed from the incubator, the old culture medium was discarded, 0.05% pancreatic enzyme was washed for 5 seconds, then sucked up, appropriate amount of 0.05% pancreatic enzyme was added, incubated at 37℃ for 2 to 5 minutes, and the cell digestion was observed under a microscope. When the cells became round and began to fall off, the digestion was terminated with primary cell termination medium, and the cells were transferred to a centrifuge tube at 1500 rpm for 3 min. The supernatant was abandoned, the cells were re-suspended in the conditioned medium of 1-5 mL, the live cells were counted, the cell suspension was inoculated on the culture bottle/plate with the appropriate cell density, and γ-NIH-3T3 cells were added (see the instructions for the area of the bottom of the culture bottle/plate with different specifications, the number of inoculated primary cells, and the number of trophoblast cells added), the conditioned medium was supplemented to the required volume, and the mixture was mixed in a cross way. 75% of the surface of the culture container was disinfected and cultured in a 5% CO2 incubator at 37 ℃. Do the same for other steps.
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Immunity
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