Human umbilical vein endothelial cell line

Immortalization of HUVEC human umbilical vein endothelial cells

Cell identification

Immunofluorescence identification has been passed

Cell source

National Resource Library (Shanghai Academy of Sciences)

Cell background

The human umbilical vein endothelial cell line HUVEC was established by ATCC in the United States a long time ago and has since been widely used. However, in recent years, it has been found that there is cross contamination at the cell source, and it has been taken down internationally. The United States has also discovered cell problems, so a new HUVEC-C has been built. However, the seeds of this cell line are in poor condition after being introduced to China and cannot be used. Therefore, it is difficult to find human umbilical vein endothelial cells (HUVE-C cells) in good condition that can be used in the market. Due to the lack of suitable cells in good and accurate condition, our company has developed a new and accurate cell line: immortalized human umbilical vein endothelial cells. Provide immunofluorescence identification of individual endothelial cell markers. The cell is called human umbilical vein endothelial cell immortalized cell, which is the same type of cell as huve-c and belongs to human umbilical vein endothelial cells. The only difference is the name, and because it is a domestically established line, STR identification cannot be provided, but immunofluorescence identification is provided.

Cell characteristics

1) Source: Human umbilical cord tissue

2) Morphology: Endothelial cell like adherent growth

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 endothelial cell culture system (recommended: PriMed-awcell-002)

Endothelial cell culture basic medium 93%

Fetal bovine serum (FBS) 5%

Endothelial cell culture additive 1%

Penicillin/Streptomycin, P/S 1%

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

L Precautions:

The human umbilical vein endothelial cell line HUVEC can refer to the following methods for passaging adherent cells:

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) trypsin 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 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. 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 subsequently subculture according to the actual situation in a ratio of 1:2~1:5.

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.

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.