104C1 (transformed guinea pig carcass cells)

104C1 (transformed guinea pig carcass cells)

All products of our company are for scientific experiments only and are not intended for use outside of scientific experiments!

Product details:

Source of organization embryo

Growth characteristics adherent cells

cell morphology Fibroblast like

Biosafety level 1

Growth medium RPMI-1640＋10% FBS＋1% P/S

Recommended passage ratio 1:3-1:4

Recommended fluid change frequency 2-3 times per week

Freezing conditions

Freezing solution:55% basic medium+40% FBS+5% DMSO

Temperature: Liquid Nitrogen

culture conditions

Gas phase: air,95%； CO2，5%

temperature37℃

Cell culture operation:

1) Resuscitate cells:The following cell culture cryopreservation treatment is for reference only, and the specific operating steps are mainly based on the accompanying product manual.

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%, subculture can be carried out.

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

b、 Add 1 mL of digestion solution (0.25% Trypsin-0.02% EDTA) to the culture bottle, allowing the digestion solution to infiltrate all cells. Place the culture bottle in a 37 ℃ incubator for 1-3 minutes (depending on the digestion status of the cells), and then observe the digestion status of the cells 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 2-3 ml of culture medium to terminate digestion. Gently mix well and transfer to a sterile centrifuge tube. Centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add 1-2 mL of culture medium, and blow well.

c、 Divide the cell suspension into a new dish or bottle containing 8 mL of culture medium at a ratio of 1:2, and place it in a culture incubator for cultivation.

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

a、 Collect cells and cell culture medium, transfer them into sterile centrifuge tubes, centrifuge at 1000 rpm for 4 minutes, discard the supernatant, wash with PBS, discard PBS, and perform cell counting.

b、 Add serum-free cell cryopreservation solution according to the number of cells, make the cell density 5 × 106~1 × 107/mL, gently mix well, and freeze 1mL of cell suspension in each cryopreservation tube, paying attention to labeling the tubes properly.

c、 Place the cryovial in a -80 ℃ freezer and transfer it to liquid nitrogen for storage after 24 hours. Record the location of the freezer for future retrieval.

Precautions for cell culture：

1After 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.

IICarefully read the cell manual to understand the relevant information of the cell, such as cell morphology, culture medium used, serum ratio, required cytokines, etc., to ensure consistent cell culture conditions. If any problems occur due to inconsistent culture conditions, the responsibility shall be borne by the customer.

IIIWipe the surface of the cell vial with 75% alcohol and observe the cell state 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.

4Adherent 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.

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

VI We suggest 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.

7This cell is for scientific research purposes only.

8Note: 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.

9Note: 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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