SCC7 (mouse squamous cell carcinoma cells)

1、 Basic cellular properties

Organizational source: Squamous cell carcinoma

Growth characteristics: Wall attached growth

Cell morphology: Epithelial like

Background introduction: SCC7 mouse squamous cell carcinoma cells refer to cells obtained from tissues or organs of the body through collagenase, other methods, and cultured in a simulated body environment in vitro.

Biosafety level: 1

Cell specifications: packaged in 1 × 106 cells/T25 culture bottles or 1mL cryovials

Culture medium: 1640+10% FBS

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

Freezing conditions: Freezing solution: 90% FBS, DMSO 10%,

Doubling time: 2 to 3 times a week

Passage ratio: 1:2 passage

Fluid change frequency: once every 2-3 days

2、 Cell culture operation

1) Resuscitation of cells: The following cell culture and cryopreservation treatments are for reference only. The specific operating steps are mainly based on the product manual provided with the goods

Quickly shake and thaw the cryovial containing 1 mL of cell suspension in a 37 ℃ water bath, and mix well with 4 mL of culture medium. Centrifuge at 1000 rpm for 3 minutes, discard the supernatant, add 1-2 mL of culture medium, and blow well. Then add all cell suspensions to a culture bottle containing an appropriate amount of culture medium and culture overnight (or add the cell suspensions to a 6 cm dish, add about 4 mL of culture medium, and culture overnight). On the third day, change the fluid and check the cell density.

2) Cell passage: If the cell density reaches 80% -90%, passage culture can be carried out.

a、 When the cells grow to cover 80% of the surface area of the culture bottle, discard the culture medium in the 25cm2 culture bottle and wash the cells once with PBS;

b、 Add approximately 1ml of 0.25% digestion solution to the culture bottle, observe under an inverted microscope, and wait for the cells to shrink and become round before adding the culture solution to terminate digestion. Gently blow the cells to detach, then transfer the suspension to a 15ml centrifuge tube and centrifuge at 1000rpm for 5 minutes;

c、 Discard the supernatant, resuspend the precipitated cells in 12ml of culture medium, and then subculture them in a 1:2 ratio. Finally, incubate the cells in a 37 ℃, 5% CO2 cell culture incubator;

d、 After the cells adhere to the wall, observe the culture results and then perform medium exchange culture or passage.

3) Cell cryopreservation: When the cells are in good growth condition, cell cryopreservation can be performed. Taking T25 bottles as an example below;

a、 When the cells grow to cover 80% of the surface area of the culture bottle, discard the culture medium in the 25cm2 culture bottle and wash the cells once with PBS;

b、 Add about 1ml of 0.25% digestion solution to the culture bottle, observe under an inverted microscope, and wait for the cells to shrink and become round before adding the culture solution to terminate digestion. Gently blow the cells to detach, then transfer the suspension to a 15ml centrifuge tube and centrifuge at 1000rpm for 5 minutes;

c、 Resuspend the cells in an appropriate amount of cryopreservation solution (FBS: DMSO=9:1) and place them in a cryovial;

d、 First, place the cell cryopreservation tube at -20 ℃ for 1.5 hours, then transfer it to -80 ℃ overnight, and after 24 hours, transfer it to liquid nitrogen for long-term storage. The program cooling box can be directly placed at -80 ℃.

3、 Cultivation precautions

1. After receiving the cells, first observe whether the cell bottle is intact and whether there is any leakage or turbidity in the culture medium. If any of the above phenomena occur, please contact us in a timely manner.

2. Carefully read the cell instructions and understand the relevant information about the cell, such as cell morphology, culture medium used, serum ratio, required cytokines, etc., to ensure consistent cell culture conditions. If there are any problems with the cell due to inconsistent culture conditions, the responsibility shall be borne by the customer.

3. Wipe the surface of the cell vial with 75% alcohol and observe the cell status under a microscope. Due to transportation issues, it is normal for some cells to form fragments due to temperature changes and severe collisions. After observing the cell state, disinfect the bottle wall with 75% alcohol and place the T25 bottle in a 37 ℃ incubator for 2-4 hours.

4. Adherent cells can be digested, suspended cells are directly mixed and collected, centrifuged at 900 rpm to 1000 rpm for 3 minutes, and the supernatant is discarded. Add 5 mL of PBS to resuspend the cells, centrifuge at 900 rpm to 1000 rpm for 3 minutes, resuspend the cells in fresh culture medium, and inoculate them into new culture bottles or dishes for cultivation in an incubator.

5. Please ask the customer to use the same culture medium for cell culture under the same conditions.

6. It is recommended that customers take several photos of the cells in the first 3 days after receiving them, recording the cell status for communication and exchange with our technical department. Due to transportation reasons, some sensitive cells may experience instability. Please contact us promptly to inform us of the specific situation of the cells, so that our technical personnel can track and follow up until the problem is resolved.

7. This cell is for scientific research purposes only.

8. Note: 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. The recommended ratio for the first passage after receiving the cells is 1:2 passage.

9. Note: 1:2 passage refers to transferring one T25 bottle to two T25 bottles or two 6cm dishes. Not one T25 bottle to two 10cm dishes.

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