KC-1317

CHOK1-B2M-FCGRT-Cell-Line

21915

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Background of CHOK1-B2M-FCGRT-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. FCGRT (Fc Fragment of IgG Receptor And Transporter) encodes a receptor that binds the Fc region of monomeric immunoglobulin G. The encoded protein transfers immunoglobulin G antibodies from mother to fetus across the placenta. This protein also binds immunoglobulin G to protect the antibody from degradation. Alternative splicing results in multiple transcript variants. Diseases associated with FCGRT include Selective Igg Deficiency Disease and Immunodeficiency 43. Gene Ontology (GO) annotations related to this gene include antigen binding and beta-2- microglobulin binding. An important paralog of this gene is HLA-C.

Specifications

Catalog Number	KC-1317
Cell Line Name	CHOK1-B2M-FCGRT-Cell-Line
Host Cell Line	Chinese hamster ovary CHO-K1 cell line
Description	CHOK1 cell line stable expressing exogenous human B2M gene and FCGRT gene
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	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+5µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 human B2M FCGRT cell line was generated using lentiviral vector expressing human B2M FCGRT sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (F12K + 10% FBS + 5µg/mL Puromycin) 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 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% F12K + 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.
Stephen A. Beers. Influence of immunoglobulin isotype on therapeutic antibody function. Blood Spotlight. 2016