KC-1081

MC38-B2M-KO-1B2 Cell Line

21501

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Background of MC38-B2M-KO-1B2 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-1081
Cell Line Name	MC38-B2M-KO-1B2 Cell Line
Host Cell Line	MC-38
Description	Stable MC38 clone with B2M gene knockout, No.1B2
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS
Selection Marker	NA
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	Yes

Cell Line Generation

MC38-B2M-KO-1B2 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of MC38-B2M-KO-1B2 cell line stable clone using PCR sequencing.

Figure 2: Characterization of MC38-B2M-KO-1B2 cell line stable clone using western blot.

Cell Resuscitation

Prewarm culture medium (DMEM + 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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

1.Wang H, Liu B, Wei J. 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. Epub 2021 Jun 12. PMID: 34129878.