KC-4431

Raji-CD19-KO-1A3-Cell-Line

46352

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Background of Raji-CD19-KO-1A3-Cell-Line

The CD19 gene, located on chromosome 16p11.2, encodes a transmembrane protein that plays a crucial role in B cell development and activation. CD19 forms a complex with CD21, CD81, and CD225, enhancing B cell receptor (BCR) signaling and lowering the threshold for B cell activation. This interaction is essential for the adaptive immune response, including the differentiation of B cells into antibody-secreting plasma cells and memory B cells. Recently, CD19 has emerged as a key target in cancer immunotherapy. Chimeric antigen receptor (CAR) T-cell therapy, which involves genetically modifying T cells to recognize and attack CD19-expressing B cells, has shown remarkable success in treating B cell malignancies such as acute lymphoblastic leukemia (ALL) and diffuse large B-cell lymphoma (DLBCL). Understanding the role of CD19 is vital for advancing immunotherapeutic strategies.

Specifications

Catalog Number	KC-4431
Cell Line Name	Raji-CD19-KO-1A3-Cell-Line
Host Cell Line	Raji
Description	Stable Raji clone with human CD19 gene knockout, No.1A3
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
Selection Marker	N/A
Morphology	Lymphoblast
Subculture	Split saturated culture 1:5-1:10 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 23 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Raji-CD19-KO-1A3 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Raji-CD19-KO-1A3 cell line stable clone using PCR sequencing.

Figure 2: Characterization of Raji-CD19-KO-1A3 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of Raji-CD19-KO-1A3 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 1:5-1:10 every 2-3 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

Carter RH, Fearon DT. CD19: lowering the threshold for antigen receptor stimulation of B lymphocytes. Science. 1992 Apr 3;256(5053):105-7. doi: 10.1126/science.1373518. PMID: 1373518. 2.https://www.ncbi.nlm.nih.gov/gene/930