KC-4432

Raji-CD19-KO-1B2-Cell-Line

46356

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Background of Raji-CD19-KO-1B2-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-4432
Cell Line Name	Raji-CD19-KO-1B2-Cell-Line
Host Cell Line	Raji
Description	Stable Raji clone with human CD19 gene knockout, No.1B2
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	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-1B2 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Raji-CD19-KO-1B2 cell line stable clone using PCR sequencing.

Figure 2: Characterization of Raji-CD19-KO-1B2 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of Raji-CD19-KO-1B2 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

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.