HEP-53.4 mouse liver cancer cells

Basic Information

Cell name：HEP-53.4 mouse liver cancer cells

Cell alias：HEP-53.4 ; 53.4 ; Mouse liver cancer cells

Cell source：Germany

Cell identification：STR identification has been passed

cell morphology：Epithelioid cells, adherent growth

Culture medium：DMEM (containing NaHCO3 1.5g/L) (BasMed-AW-013)+FBS 10%+P/S1%

culture conditions：Gas phase: Air, 95%; Carbon dioxide, 5%. Temperature: 37 degrees Celsius, humidity in the incubator is 70% -80%

Freezing conditions：Serum free cryopreservation solution, stored in liquid nitrogen

Cellular background：Primary hepatocellular carcinoma originating from C57BL/6J mice, these mouse liver cells faithfully represent hepatocellular carcinoma and provide valuable models for studying this type of liver cancer, synonymous with HEP-53.4 and 53.4, which are easy to identify and cross reference. Hepatocellular carcinoma is a major health issue, and these cells can be accurately studied for their molecular pathways, cell interactions, and treatment strategies. These cells originate from Mus musculus (mice) and provide a relevant model system for understanding and developing treatment methods for hepatocellular carcinoma.

Cell usage：For scientific research purposes only

Cell delivery time: In stock, about 1 week

Precautions

1. Conventional digestion, collection of cells, and centrifugation.

2. After centrifugation, remove the supernatant from the centrifuge tube, add about 1ml of resuspended cells and mix well. It is recommended to gently shake or blow the cells, put them into the incubator for digestion, and then digest for about 1 minute.

3. After digestion, gently blow the cell suspension with a pipette to disperse the cell clusters. Quickly add 3-5ml of serum containing culture medium and mix well to terminate digestion. Centrifuge to remove.

4. Add about 5ml of the corresponding culture medium to the cells and mix well. Transfer the mixture into the culture bottle/dish in proportion.

5. Observe under a microscope to see if the cells are evenly dispersed as single cells. If there are a small number of clustered small cell clusters, they do not need to be digested again. Let them adhere to the wall and wait for the cells to grow stably before dissipating.

Shipping at room temperature

After receiving the T25 bottle, disinfect it and place it in the incubator for 2-3 hours. Observe the density and condition, take 2-3 photos, and provide feedback to the sales team. Once the density meets the standard, it can be passaged. The initial passage ratio is 1:2. It is recommended to freeze one whole bottle into a 1ml cryovial and continue passaging the other bottle. Repeat the freezing process for 2-3 bottles before amplification for experimentation to prevent seed breakage in case of unexpected situations.

Dry ice shipmentTwo conventional cell shipping cryovials were used, one was revived and the other was kept as a backup. If the first one failed to revive, the second one was resuscitated strictly according to the manufacturer's requirements. If none of the resuscitations were successful, please keep a photo of the resuscitation and notify us immediately.

HEP-53.4 mouse liver cancer cells

adherent cells

1. Discard the culture supernatant and rinse the cells 1-2 times with PBS that does not contain calcium or magnesium ions.

2. Add 0.25% (w/v) 0.53 mM EDTA to culture bottles (1-2mL for T25 bottles and 2-3mL for T75 bottles), digest at 37 ° C for 1-2 minutes (digestion time can be appropriately extended for difficult to digest cells), and then observe the cell digestion under a microscope. If most of the cells become round and fall off, quickly take them back to the operating table, tap the culture bottle a few times, and add 3-4ml of culture medium containing 10% FBS to terminate digestion.

3. Gently mix and aspirate, centrifuge at 1000RPM for 3-5 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly. Divide the cell suspension into new T25 bottles/6cm culture dishes in a ratio of 1:2, add 6-8ml of the new culture medium prepared according to the instructions to maintain the growth vitality of the cells, and subsequently subculture according to the actual situation in a ratio of 1:2~1:5.

4. Cell cryopreservation: After receiving the cells, it is recommended to freeze a batch of cell seeds during the first 3 generations of cultivation for subsequent experiments.

5. The transport medium (infusion medium) cannot be used to culture cells anymore. Please use a newly prepared medium according to the instructions for cell culture conditions to culture cells.

Suspended cells

Cells grown in suspension can be maintained in their growth state by adding culture medium to the culture bottle. Generally, maintaining a cell density of 1 × 10 ⁵~1 × 10 ⁶ cells/mL (different cells have different density requirements) can maintain normal cell growth. If necessary, the cell suspension can be collected in a centrifuge tube at 1000rpm and centrifuged for 5 minutes. Discard the supernatant, add 1-2mL of culture medium, resuspend and mix well. Then, divide the cell suspension into new T25 bottles at a ratio of 1:2 and add 6-8ml of new culture medium prepared according to the instructions to maintain the growth vitality of the cells. Subsequent passages should be carried out at a ratio of 1:2-1:4 according to the actual situation.

mode of transportation

Low temperature: Transport in 1mL cryovials packaged with dry ice. After receiving, store overnight in a -80 degree freezer and transfer to liquid nitrogen or directly resuscitate. If you find that the dry ice has evaporated completely, the cryovial cap has fallen off, is damaged, or the cells are contaminated, please contact us immediately.

At room temperature: T25 bottles of revived surviving cells are shipped at room temperature and processed according to the procedures for cell reception upon receipt.

biosafety

1. All animal 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 before disposal.

2. It is recommended to always use protective gloves, clothing, and a face mask when reviving frozen cells. Attention: The cryotube immersed in liquid nitrogen will leak and gradually fill with liquid nitrogen. When thawing, the conversion of liquid nitrogen into gas phase may cause the container to explode or the lid to be blown off with dangerous force, resulting in flying debris and causing personal injury.

Special attention

The cell grows well in DMEM (containing 1.5g/L NaHCO3) medium, most of which contain high concentrations of NaHCO3 (3.7g/L). If DMEM (3.7g/L NaHCO3) medium is used to culture cells, the CO2 concentration needs to be increased (7% -10%).