3T3-L1 mouse embryonic fibroblast brand

Product information:

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

Temperature: Liquid Nitrogen

Training Plan A (default)：Growth medium: DMEM+10% NCS+1% P/S

Cultivation conditions:Gas phase: Air, 95%; CO2， 5%, temperature: 37 ℃

Recommended passage ratio1:3-1:4

Background description3T3-L1 cells are consecutive sub strains of 3T3 (swiss mice) isolated through cloning. When 3T3-L1 cells transition from rapid division to full growth and contact inhibition, 3T3-L1 cells undergo preadipolar to adipoid reversal. In the culture medium, high serum content can promote the accumulation of intracellular fat in 3T3-L1 cells.

Age (gender)embryo

cell typeSpontaneous immortalized cells

Biosafety levelBSL-1

Receptor expression statusinsulin, expressed

Preservation institutionATCC; CL-173 ATCC; CCL-92.1BCRC; 6015

Transportation and storage:

Dry ice transportation and recovery of viable cells

(1) 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.

(2) T25 bottles of revived surviving cells will be shipped at room temperature and processed according to the procedures for cell reception upon receipt.

Processing after cell reception:

1) After receiving the cells, disinfect the bottle wall with 75% alcohol and place the T25 bottle in a 37 ℃ incubator for about 2-3 hours. If you find any damage to the culture bottle, overflow of liquid, or contamination of the cells, please take photos and contact us promptly.

2) Please confirm the cell status under a 4 or 5X microscope, and take 2-3 photos (10 ×, 20 ×) of the newly received cells, as well as one photo of the appearance of the culture bottle, for retention as a basis for the cell status upon receipt during after-sales service.

3) Adherent cells: Cells are placed in a 37 ℃ incubator for 2-3 hours, and their growth and adherence are observed under a microscope. Some adherent cells may detach and form clusters due to vibration during express delivery. If the growth density of cells observed under the microscope is below 60%, the culture medium in the culture bottle can be removed (if there are unattached cells, they need to be recovered by centrifugation and resuspended into the original culture bottle), and 6-8mL of newly prepared culture medium can be added and placed in the cell culture box for further cultivation. If the cell growth density reaches 70% -80% or more, the cells can be passaged. During the passage process, if cells shed due to transportation vibrations, they need to be recovered by centrifugation.

Cell processing:

1) Recovery of frozen cells:

Quickly shake and thaw the cryovial containing 1mL of cell suspension in a 37 ℃ water bath, then add it to a centrifuge tube containing 4-6mL of culture medium and mix well. Centrifuge at 1000RPM for 3-5 minutes, discard the supernatant, and resuspend the cells in the culture medium. Then add the cell suspension to a culture bottle (or dish) containing 6-8ml of culture medium and culture overnight at 37 ℃. The next day, observe cell growth and cell density under a microscope.

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

For the passage of adherent cells, the following methods can be referred to:

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 in a ratio of 1:2, add 6-8ml of new culture medium prepared according to the instructions to maintain the growth vitality of the cells, and proceed with subsequent passages in a ratio of 1:2 to 1:5 based on the actual situation.

3) Cell cryopreservation: It is recommended to freeze a batch of cell seeds during the first 3 generations of culture after receiving the cells for subsequent experimental use.

The products currently being sold by the company: