Human endometrial cancer cell line EFE184

Human endometrial cancer cellsEFE184

Introduction:The EFE184 cell line was established from ascites of a 69 year old woman with recurrent endometrial cancer. The cell morphology is a single-layer epithelioid cell, and its growth is very slow. The EFE184 cell line can be used for 3D cell culture and cancer research.

Item Number：WS-S1480

specification：1 × 10 ⁶ cells/T25 culture bottle

1、 Basic cellular information

-Cell source: In situ: uterus; Transfer site: Ascites

-Cell morphology: Epithelial like, adherent growth

-Cell content:>1 × 10 ⁶ cells/bottle

-Packaging specificationsT25 culture bottle/1mL cryovial

-Product Usage: For scientific research purposes only,Cannot be used as a therapeutic product for animal or human diseases.

2、 Culture medium and conditions

1. Culture medium

RPMI1640 medium; 10% fetal bovine serum; 1% dual antibody

Recommended serum:WinSera FBS500 - WP - 002

2. Cultivation conditions

-Gas phase: 95% air+5% CO ₂ Temperature: 37 ℃; Incubator humidity: 70% -80%

3. Freezing and storage

-Cryopreservation solution: 90% fetal bovine serum+10% DMSO (currently prepared and used)

-Storage conditions: Long term storage in liquid nitrogen

3、 Transportation form and receiving processing

1. Shipping at room temperature (T25 bottles of live cells)

After receiving the cells, first Surface disinfection of T25 culture bottle body; Place the culture bottle in the incubator and let it stand for 2-3 hours; Observe cell density and growth status, take 2-3 clear photos and provide feedback to sales; Once the cell density meets the standard, passaging can proceed. The recommended ratio for early passaging is 1:2; After the cells are fully grown again, freeze one whole bottle of cells into one 1mL cryovial, and continue passaging the other bottle; It is recommended to repeatedly freeze 2-3 seed cells before conducting expansion experiments to avoid cell breakage due to unexpected situations.

Note: The transport medium (infusion medium) cannot be used for cell culture anymore. Please use a newly prepared medium according to the instructions for cell culture conditions to culture the cells. Suggestions for subculturing after receiving cells T25 culture bottles are passaged at a ratio of 1:2.

2. Dry ice shipment (frozen tube cells)

Regular shipping is Two cryovials, it is recommended to revive one and keep one as a backup; If the recovery fails, strictly follow the manufacturer's standard recovery process to operate the second one; If both devices fail to recover, please immediately retain photos of the entire recovery process and notify sales for handling.

4、 Cell culture steps

1. Recovery of frozen cells

-Place a 1mL cell suspension cryovial in a 37 ℃ water bath and shake rapidly until thawed;

-Gently mix in a centrifuge tube containing 4-6 mL of culture medium;

-Centrifuge at 1000RPM for 3-5 minutes and discard the supernatant;

-Resuspend cells in culture medium and inoculate them into culture bottles/dishes containing 6-8 mL of culture medium;

-Cultivate overnight at 37 ℃, and observe the cell growth status and density under a microscope the next day.

2. For passaging of adherent cells, the following methods can be referred to: (Cell density of 80% -90% can be passaged)

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

-Add 0.25% (w/v) yi protease-0.53 mM EDTA to culture bottles (1-2mL for T25 bottles and 2-3mL for T75 bottles), digest in a 37 ℃ incubator 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 workstation, tap the culture bottles a few times, and add 3-4ml of culture medium containing 10% FBS to terminate digestion.

-Gently mix and aspirate, centrifuge at 1000RPM for 3-5 minutes, discard the supernatant, add 1-2 mL 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 1:2-1:5 according to the actual situation.

For suspended cells, the following methods can be used for passaging:

-Note: The recommended passage ratio for the second passage is 1:2, and the passage ratio in the future can be determined according to customer needs.

-Method 1: Collect cells, centrifuge at 1000RPM for 4 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly. Divide the cell suspension into new dishes or bottles containing 8ml of culture medium in a ratio of 1:2 to 1:5.

-Method 2: You can choose the method of half changing the medium. After discarding half of the medium, suspend the remaining cells and divide the cell suspension into new dishes or bottles containing 8ml of medium in a ratio of 1:2 to 1:3.

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、 Biosafety precautions

1. All animal cells are considered to have potential biological hazards and must be operated in an ER level biosafety cabinet with proper personal protection; All waste liquids and containers that have come into contact with cells must be sterilized before disposal.

When reviving frozen cells, it is necessary to wear protective gloves, protective fu, and anti hu face shields; Immersing cryovials in liquid nitrogen may result in leakage and the infusion of liquid nitrogen. During thawing, the liquid nitrogen may vaporize, leading to container rupture, bottle cap breakage, and personal injury such as scratches. Caution must be exercised during operation.