KC-0962

MC38-mB2M-KO-1B3-Cell-Line

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Background of MC38-mB2M-KO-1B3-Cell-Line

B2M, also known as β2 microglobulin, is a component of MHC class I molecules and is expressed on nearly all nucleated cells. B2M noncovalently lies beside α3 heavy chain of MHC class I molecules on the cell surface without transmembrane region.

Specifications

Catalog Number	KC-0962
Cell Line Name	MC38-mB2M-KO-1B3-Cell-Line
Host Cell Line	Mouse MC38 cell line
Description	MC38 cell line with B2M gene knockout
Quantity	One vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS
Morphology	Epithelial
Subculture	Split saturated culture 1:2-1:5 every 3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 48 hours
Mycoplasma Status	Negative
In Vivo Validation	Yes

Cell Line Generation

MC38-B2M knockout cell line was generated using the CRISPR method.

Characterization

Figure: Characterization of B2M knockout in MC38 using PCR sequencing.

Figure 2: Characterization of B2M knockout in MC38 using western blot.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:2-1:5 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Hanbing Wang, Baorui Liu, Jia Wei ,Beta2-microglobulin(B2M) in cancer immunotherapies: Biological function, resistance and remedy. Cancer Lett. 2021 Oct 1;517:96-104. doi: 10.1016/j.canlet.2021.06.008.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.