KC-5216-DW

K562-BCMA-CD19-GFP-Luc2 Cell Line

60267

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Background of K562-BCMA-CD19-GFP-Luc2 Cell Line

CD19 is a B-lineage-specific transmembrane glycoprotein, the expression of which is maintained on more than 95% B-cell malignancies. This strict lineage restriction makes CD19 an ideal target for immune therapies using chimeric antigen receptors (CARs).CD19 is nearly ubiquitously expressed on B-lymphocytes and in B-cell malignancies. The anti-CD19 monoclonal antibody tafasitamab, paired with the immunomodulator lenalidomide, mediates antibody-dependent cellular toxicity and phagocytosis; the antibody-drug conjugate loncastuximab tesirine delivers the DNA cross-linking agent tesirine via CD19 binding and internalization; and CD19-directed chimeric antigen receptor T-cell therapy (CAR-T) products are engineered from autologous T cells.
B cell maturation protein (BCMA), also named CD269, is a membrane of the TNF receptor super family (TNFRSF17) and mainly expressed in mature B-lymphocytes, and play an important role in B cell development and autoimmune response.

Specifications

Catalog Number	KC-5216-DW
Cell Line Name	K562-BCMA-CD19-GFP-Luc2 Cell Line
NCBI/UniProt Accession Number	NM_001770.5, NM_001192.3
Clone Number	8#
Host Cell Line	K562-GFP-Luc2
Description	Stable K562-GFP-Luc2 cell line expressing exogenous BCMA and CD19 gene
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 + 1μg/mL Puromycin + 300μg/mL Hygromycin B
Selection Marker	Puromycin \| Hygromycin B
Morphology	lymphoblast
Subculture	Split saturated culture 1:2-1:4 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

K562-BCMA-CD19-GFP-Luc2 Cell Line was generated using a lentiviral vector expressing the human CD19 and BCMA sequence.

Characterization

Figure1: Characterization of human CD19 and BCMA overexpression in K562-BCMA-CD19-GFP-Luc2 stable clones using FACS.

Figure2: Characterization of the K562-BCMA-CD19-GFP-Luc2 cell line stable clone using PCR.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 1μg/mL Puromycin and 300μg/mL Hygromycin B)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:2-1:4 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage.

References

1. Sermer D, Elavalakanar P, Abramson JS, Palomba ML, Salles G, Arnason J. Targeting CD19 for diffuse large B cell lymphoma in the era of CARs: Other modes of transportation. Blood Rev. 2023 Jan;57:101002.
2. Hatzoglou, A. et al. TNF Receptor Family Member BCMA (B Cell Maturation) Associates with TNF ReceptorAssociated Factor (TRAF) 1, TRAF2, and TRAF3 and Activates NF- B, Elk-1, c-Jun N-Terminal Kinase, and p38 Mitogen-Activated Protein Kinase. The Journal of Immunology 165, 1322ÿ1330 (2000).

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.