Mouse fibroblast cells

Cell identification

Species identification has been passed

Cell source

National Resource Library

Cell background

In March 1948, NCTC clone 929 (mouse connective tissue) was established as a clone of cell line L. Cell line L is one of the earliest established continuous culture cell lines, and clone 929 is the earliest cloned strain. A parental cell line L was established by inserting normal subcutaneous loose connective tissue from a 100 day old male C3H/An mouse into adipose tissue. Clone929 was established by isolating single cells from the 95th generation cell line L using capillary method. The test found that the mouse pox virus was negative.

Cell characteristics

1) Source: Organization: Subcutaneous connective tissue; Loose connective tissue and fat strain: C3H/An;

2) Morphology: Fibroblasts, adherent growth;

3) Content:>1x106 cells;

4) Specification: packaged in T25 bottles or 1mL cryovials;

5) Purpose: For scientific research purposes only.

Cultivation conditions

1) Prepare MEM (recommended awcell-0012) culture medium; Horse serum (recommended: awcell-002b) 10%; Double antibody, 1%. This cell cannot be cultured with fetal bovine serum.

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

L Precautions

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) 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 form

Low temperature: (1) Transport 1mL frozen storage tube packaged with dry ice. After receiving it, store it in a -80 degree freezer overnight and transfer it to liquid nitrogen or directly recover it. If the dry ice has evaporated completely, the bottle cap of the frozen storage tube has fallen off, is damaged, and 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.