T98G-LUC human pleomorphic malignant glioma - luciferase labeling

Characteristics:

Safety: All tumor and virus transfected cells are considered to have potential biological hazards and must be operated in a secondary biosafety platform. Please pay attention to protection, and all waste liquids and containers that have come into contact with these cells must be sterilized under high pressure before disposal. After receiving the cells, under an inverted microscope*Okay, there it is4Xobjective lens)Observe the overall growth of the cells.

（1） If the cells are not fully grown, use75%After disinfecting the entire bottle with alcohol spray, place it in the super sterilization table and perform strict aseptic operation. Open the cell culture bottle, aspirate the culture medium, and replace it10mlContinue cultivation after fresh culture medium.(two)If the cells are already fully grown, they can be passaged and cultured. The specific steps are as follows:

1. Discard the culture medium and usePBS(Calcium free),Magnesium ion washing1-2Next time.

2. plus1mlDigestive fluid（0.25%Trypsin-0.53mM EDTA）In the culture bottle, pour it upside down37℃ incubator1-3Preheat for minutes, then flip the culture bottle over10-30After a few seconds, observe the digestion of cells under an inverted microscope. If most of the cells become round, quickly take them back to the operating table, tap the culture bottle a few times, and add a small amount of * culture medium to terminate digestion.

3. press6-8ml/Add * culture medium to the bottle, gently mix well and aspirate half, then transfer to a new culture bottle. If not otherwise specified, after receiving the cells*The second generation is usually one to two.

Notes:

1Observe cells under low magnification（4or5Xobjective lens)Proceed below, otherwise the passaging density of cells cannot be accurately determined. Please observe the morphology of the cells10Xor20XUnder the objective lens.

2The culture medium transported in bottles cannot be reused,Please switch to a new culture medium with dual antibodies and freeze the cells,No antibiotics can be added to the culture medium.

3Some cells are not firmly attached to the wall, and if they are found to have fallen off, they can be centrifuged and blown before being inoculated into a new bottle.

4After receiving the cells, if you find that the culture bottle is damaged, there is liquid overflow, or the cells are contaminated, please contact us promptly.