Human glioma cell U373

Human glioma cellsU373

Introduction:Problematic cell line: Misidentified/contaminated. Shown to be a U-251MG derivative. This cell line is not the original astrocytoma cell line established in 1973 at the University of Uppsala. See U-373MG Uppsala (Cellosaurus=CVCL_2818) for the original U-373MG cell line。

Item Number：WS-S1465

specification：1 × 10 ⁶ cells/T25 culture bottle

1、 Basic cellular information

-Cell source: Brain

-Cell morphology: Epithelial like, adherent growth

-Cell content:>1 × 10 ⁶ cells/bottle

-Packaging specifications: T25 culture bottle/1mL cryovial

-Product Usage: For scientific research purposes only,Cannot be used as a therapeutic product for animal or human diseases.

II Detailed rules for handling cells at room temperature on the day of receipt

Step: Receipt and Inspection

·The first and second steps of the execution section (general process).

Step 2: Change the culture medium and collect cells

·Behind the disinfectant bottle, open the bottle cap in the ultra clean workbench.

·Replace with the culture medium provided with the product.

·Special attention: If a large number of cells are found suspended under the microscope, transfer all the culture medium in the bottle to a sterile centrifuge tube, centrifuge (such as 1200rpm, 5 minutes) to collect the cells, and discard the old culture medium.

·After completing the replacement or collection, place the culture bottle in the incubator and let it sit for another 2-3 hours.

Step 3: Post treatment after standing (based on cell density)

III Steps for Cell Passage Operation

A. Subculture of adherent cells

1. Preparation: Discard the old culture medium.

2. Rinse: Slowly add washing solution such as PBS along the side wall of the culture bottle, gently shake and then aspirate to remove residual serum.

3. Digestion: Add preheated enzyme (such as T25 plus 1ml), gently shake to cover all cells with enzyme.

4. Incubation: Digest at room temperature for about 2 minutes (time varies by cell).

5. Observation: Observe under a microscope until ≥ 90% of cells become round and shed. If not achieved, check every 30 seconds.

6. Termination: Once the cells have fully dissociated, immediately add twice the volume of enzyme to the preheated culture medium to terminate digestion.

7. Blowing: Gently blow the bottle wall with a pipette to allow the cells to detach and disperse into a single-cell suspension.

8. Centrifuge: Transfer the cell suspension to a centrifuge tube, centrifuge at 200 × g for 3-5 minutes, and discard the supernatant.

9. Resuspend and Inoculate: Resuspend the cell pellet with a small amount of fresh culture medium, dilute it in an appropriate proportion, and inoculate it into a new culture bottle.

10. Cultivation: Place in the incubator. If using a regular bottle cap, be sure to loosen the cap to facilitate gas exchange.

B. Suspension cell passage

1. Collection and centrifugation: Transfer all cell suspensions to centrifuge tubes, centrifuge at 1000 rpm for 5 minutes, and discard the supernatant.

2. Resuspend: Add 1-2ml of fresh culture medium and gently resuspend the cells.

3. Inoculation: Transfer the cell suspension to a new T25 culture bottle in a ratio of 1:2, and supplement the total amount of culture medium to 5-8ml.

4. Cultivation: Place it in the incubator for further cultivation.

4 Special problem handling: adherent cells detached during transportation

Some cells that are not firmly attached to the wall will fall off during transportation, which is a normal phenomenon. Follow the steps below to handle it:

1. Collection: Transfer all the culture medium in the bottle into a sterile centrifuge tube, centrifuge (1200rpm, 3-5 minutes), and discard the supernatant.

2. Cleaning: Resuspend the cells in PBS, centrifuge again (1200 rpm, 3-5 minutes), and discard the PBS.

3. Digestion: Add about 1ml of 0.25% enzyme, gently mix (do not vigorously blow), and put it in the incubator for 1-2 minutes for digestion.

4. Termination and dispersion: After removal, gently blow to disperse the cell clusters, quickly add 3-5ml of culture medium to terminate digestion, centrifuge (1200rpm, 3-5 minutes) and discard the supernatant.

5. Resuspend and Inoculate: Resuspend with 5ml of culture medium and inoculate into new culture bottles/dishes in proportion.

6. Observation: Observe under a microscope. If the cells are single or small clusters of only 3-5 cells, they can be cultured normally and allowed to adhere to the wall for growth. --

5 Summary of After sales Service Policy

Important NoticeAny abnormal issues,Take photos of timeAnd contacting us is the key to obtaining after-sales support.