M2-10B4细胞株小鼠骨髓纤维原细胞系-NTCC保藏中心

M2-10B4 Cell Line

Cat No.: BioVector-A1972

Product category

Animal cells

Organism

Mus musculus, mouse

Classification

Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod

Morphology

fibroblast

Tissue

Bone; Marrow; Stroma

Applications

3D cell culture

Immunology

Characteristics

Growth properties

Adherent

Derivation

The M2-10B4 cell line is a clone derived from bone marrow stromal cells from a (C57BL/6J X C3H/HeJ)F1 mouse.

Genes expressed

laminin; collagen IV

Comments

The M2-10B4 cell line is a clone derived from bone marrow stromal cells from a (C57BL/6J X C3H/HeJ)F1 mouse.

The cells express laminin and collagen IV, but do not express collagen I or Factor VIII.

The cells will support human and murine myelopoiesis in long term culture, and will support certain murine stromal cell dependent pre-B cell lines in vitro.

The base medium for this cell line is ATCC-formulated RPMI-1640 Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum  to a final concentration of 10%.

Handling information

Unpacking and storage instructions

1. Check all containers for leakage or breakage.

2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until ready for use.

Complete medium

The base medium for this cell line is ATCC-formulated RPMI-1640 Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum  to a final concentration of 10%.

Temperature

37°C

Atmosphere

95% Air, 5% CO2

Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).

2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

3. Transfer the vial contents to a 75 cm2 tissue culture flask and dilute with the recommended complete culture medium (see the specific batch information for the recommended dilution ratio).   It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).

4. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

If it is desired that the cryoprotective agent be removed immediately, or that a more concentrated cell suspension be obtained, centrifuge the cell suspension at approximately 125 xg for 5 to 10 minutes.  Discard the supernatant and resuspend the cells with fresh growth medium at the dilution ratio recommended in the specific batch information.

Subculturing procedure

Remove medium, and rinse with 0.25% trypsin, 0.53 mM EDTA solution. Remove the solution and add an additional 1 to 2 mL of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks.

Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended

Medium Renewal: Every 2 to 3 days

Reagents for cryopreservation

Complete growth medium supplemented with 5% (v/v) DMSO

BSL: Level 1

Supplier: BioVector NTCC Inc.

www.biovector.net