C57BL/6J小鼠肝细胞癌细胞系Hep-53.4

Organism
物种

Mouse小鼠

Tissue
组织

Liver肝脏

Disease
疾病

Hepatocellular   carcinoma小鼠肝细胞癌

Synonyms
同名词

HEP-53.4, 53.4

Age
年龄段

Adult成年

Gender
性别

Female 雌性

Morphology
细胞形态

Epithelial-like   类上皮细胞

Growth   properties
生长特性

Adherent 贴壁细胞

Citation

Hep-53.4   (Cytion 货号 400200)

Biosafety level

1

Tumorigenic

Yes, in   C57BL/6J mice

Mutational   profile

p53 wt

Culture Medium

DMEM, w: 4.5   g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium   pyruvate (Cytion article number 820300a)

Medium   supplements

Supplement the   medium with 10% FBS

Passaging   solution

Accutase

Subculturing

Remove the old   medium from the adherent cells and wash them with PBS that lacks calcium and   magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10   ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25   flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature   for 8-10 minutes to detach them. After incubation, gently mix the cells with   10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes.   Discard the supernatant, resuspend the cells in fresh medium, and transfer   them into new flasks that already contain fresh medium.

Split ratio

A ratio of 1:4   to 1:8 is recommended

Fluid renewal

Every 3 to 5   days

Freeze medium

CM-1 (Cytion   catalog number 800100) or CM-ACF (Cytion catalog number 806100)

Handling of   cryopreserved cultures

1. Confirm that the vial remains deeply frozen upon       delivery, as cells are shipped on dry ice to maintain optimal       temperatures during transit.

2. Upon receipt, either store the cryovial       immediately at temperatures below -150°C to ensure the preservation of       cellular integrity, or proceed to step 3 if immediate culturing is       required.

3. For immediate culturing, swiftly thaw the vial by       immersing it in a 37°C water bath with clean water and an antimicrobial       agent, agitating gently for 40-60 seconds until a small ice clump       remains.

4. Perform all subsequent steps under sterile       conditions in a flow hood, disinfecting the cryovial with 70% ethanol       before opening.

5. Carefully open the disinfected vial and transfer       the cell suspension into a 15 ml centrifuge tube containing 8 ml of       room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to       separate the cells and carefully discard the supernatant containing       residual freezing medium. Optionally, skip centrifugation but remove any       remaining freezing medium after 24 hours.

7. Gently resuspend the cell pellet in 10 ml of fresh       culture medium. For adherent cells, divide the suspension between two       T25 culture flasks; for suspension cultures, transfer all the medium       into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for       continued growth and maintenance of the cell line, ensuring reliable       experimental outcomes.

Sterility

Mycoplasma   contamination is excluded using both PCR-based assays and luminescence-based   mycoplasma detection methods.

To ensure there   is no bacterial, fungal, or yeast contamination, cell cultures are subjected   to daily visual inspections.

STR profile

M_18-3: 16

M_4-2: 20.3

M_6-7: 17

M_3-2: 14

M_19-2: 13

M_7-1:   26.2,27.2

M_1-1: 16

M_8-1: 16

M_2-1: 15

M_15-3: 22.3

M_6-4: 18

M_11-2: 16

M_1-2: 20

M_17-2: 15

M_12-1: 17

M_5-5: 16,17

M_X-1: 28

M_13-1: 17

Human D4/D8: -