KC-5419

NCI-H929-BCMA-KO Cell Line

56411

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Background of NCI-H929-BCMA-KO Cell Line

BCMA, i.e., CD269 or TNFRSF17, is a TNF receptor superfamily 17 member, expressed on differentiated plasma cells and plasmablasts under physiological conditions and nearly on all MM tumor cells. BCMA ligands include APRIL (A Proliferations-Inducing Ligand) and BAFF (B-cell activating factor) which are involved in the maturation and differentiation of PCs. APRIL can bind to BCMA more avidly than to BAFF, and both can induce BCMA downstream signals to PI3K-PKB/Akt (i.e., phosphoinositide-3-kinase-protein kinase B/Akt), to RAS/MAPK (i.e., rat sarcoma/mitogen-activated protein kinase), and also to NF-κB (i.e., nuclear factor kappa-B), inducing increased plasma cells proliferation and survival.

Specifications

Catalog Number	KC-5419
Cell Line Name	NCI-H929-BCMA-KO Cell Line
Clone Number	5C2
Host Cell Line	NCI-H929
Description	Stable NCI-H929 clone with human BCMA gene knockout, No.5C2
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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+0.05mM 2-mercaptoethanol
Selection Marker	NA
Morphology	lymphoblast
Subculture	Split saturated culture 1:5-1:7 every 2-3 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 50 hours
Mycoplasma Status	Negative

Cell Line Generation

NCI-H929-BCMA-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of NCI-H929-BCMA-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of NCI-H929-BCMA-KO cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of NCI-H929-BCMA-KO cell line stable clone using FACS.

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 1:5-1:7 every 2-3 days; seed out at about 1-3 ×10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Tan CR, Shah UA. Targeting BCMA in Multiple Myeloma. Curr Hematol Malig Rep. 2021 Oct;16(5):367-383. doi: 10.1007/s11899-021-00639-z. Epub 2021 Aug 25. PMID: 34432234; PMCID: PMC9655650.
Sammartano V, Franceschini M, Fredducci S, Caroni F, Ciofini S, Pacelli P, Bocchia M, Gozzetti A. Anti-BCMA novel therapies for multiple myeloma. Cancer Drug Resist. 2023 Mar 22;6(1):169-181. doi: 10.20517/cdr.2022.138. PMID: 37065871; PMCID: PMC10099603.