Human embryonic kidney cell 293T

HEK-293T human embryonic kidney cells

HEK-293T‘293T’.

Cell identification

STR identification has been passed

Cell source

National Resource Library

Cell background

HEK-293T cells are derivatives of the 293T (293tsA1609neo) cell line (ATCC CRL-11268). The cells continuously express SV40 antigen, which is commonly used in transfection experiments with high transfection efficiency. (Transfection is usually performed at a density of 50% on the plate, and after the cells have just attached to the dish wall (usually 8-10 hours), transfection can be carried out.)

Cell characteristics

1) Source: Human embryonic kidney

2) Morphology: Epithelial like, adherent growth, weak adhesion ability

3) Content:>1x106 cell count

4) Specification: Packaging in T25 bottles or 1mL cryovials

5) Purpose: For scientific research purposes only

Cultivation conditions:

1) Prepare DMEM (recommended awcell-0001) culture medium; High quality fetal bovine serum, 10%; L-glutamine (2mM) 1%; NEAA 1% ； Double antibody 1%

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

L Precautions:

This cell is a suspended and slightly adherent cell, and passaging can refer to the following methods:

1. Collection: Collect the suspended cells in the culture bottle into a centrifuge tube. Wash cells 1-2 times with PBS that does not contain calcium or magnesium ions. Due to the weak adhesion of cells to the wall, cells will fall off after PBS washing, so PBS also needs to be recovered into centrifuge tubes.

2. Add 0.25% (w/v) trypsin 0.53 mM EDTA to culture bottles (T25 bottle 1-2mL, T75 bottle 2-3mL) and digest in a 37 ° C incubator for 1-2 minutes (difficult to digest cells can be appropriately prolonged for digestion time). Then observe the digestion of 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 3-4ml of culture medium containing 10 μ S to terminate digestion.

3. Centrifuge the collected suspended cells, cells in PBS cleaning solution, and digested adherent cells at 1000rpm for 5 minutes, discard the supernatant, add 1-2mL of culture medium, resuspend and mix well, and then divide the cell suspension into new T25 bottles at 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. Subsequent passages should be carried out according to the actual situation at a ratio of 1:2~1:5.

Cryopreservation of human embryonic kidney cell 293T cells: After receiving the cells, it is recommended to freeze a batch of cell seeds during the first 3 generations of culture for subsequent experiments.

Transportation mode:

Low temperature: (1) Transport in 1mL cryovials packaged with dry ice. After receiving, store in a -80 degree freezer overnight 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.

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

Biosafety:

1. All animal cells are considered to have potential biological hazards and must be operated in a secondary biosafety platform. Please pay attention to protection, and all waste liquids and containers that have come into contact with these cells must be sterilized before disposal.

2. It is recommended to always use protective gloves, clothing, and a face mask when reviving frozen cells. Attention: The cryotube immersed in liquid nitrogen will leak and gradually fill with liquid nitrogen. When thawing, the conversion of liquid nitrogen into gas phase may cause the container to explode or the lid to be blown off with dangerous force, resulting in flying debris and causing personal injury.