KC-3882

CHOK1-mouse-CD19-Cell-Line

34960

Home » 细胞系 » CHOK1-mouse-CD19-Cell-Line

Background of CHOK1-mouse-CD19-Cell-Line

CD19 encodes a member of the immunoglobulin gene superfamily. The expression of this cell surface protein is limited to B cell lymphocytes. This protein is a reliable biomarker for pre-B cells, but its expression decreases during the differentiation of terminal B cells in plasma cells secreting antibodies. This protein forms complexes with various membrane proteins, including type 2 complement receptor (CD21) and transmembrane tetraprotein (CD81), and this complex reduces the threshold for antigen induced B cell activation. The activation of this B-cell antigen receptor complex activates the phosphatidylinositol 3-kinase signaling pathway, followed by the release of intracellular calcium ions. This protein is a target of chimeric antigen receptor (CAR) T cells used for the treatment of lymphocytic leukemia. The mutation of this gene is associated with the common variant immunodeficiency disease 3 (CVID3), which leads to failure of B cell differentiation and impaired immunoglobulin secretion. The characteristics of CVID3 are hypogammaglobulinemia, inability to produce antibody responses to antigens, and recurrent bacterial infections.

Specifications

Catalog Number	KC-3882
Cell Line Name	CHOK1-mouse-CD19-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous mouse-CD19 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% RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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
In Vivo Validation	NA

Cell Line Generation

CHOK1-mouse-CD19-cell-line was generated using a lentiviral vector expressing the mouse-CD19 sequence.

Characterization

Figure 1: Characterization of mouse-CD19 overexpression in the CHOK1-mouse-CD19 stable clone using FACS..

Figure 2: Characterization of mouse-CD19 in the CHOK1-mouse-CD19 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/mL puromycin)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 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI 1640 + 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.Francis J, Dharmadhikari AV, Sait SN, Deeb G, Wallace PK, Thompson JE, Wang ES, Wetzler M. CD19 expression in acute leukemia is not restricted to the cytogenetically aberrant populations. Leuk Lymphoma. 2013 Jul;54(7):1517-20. doi: 10.3109/10428194.2012.754096. Epub 2013 Jan 3. PMID: 23193950; PMCID: PMC6668030. 2.Awan FT, Lapalombella R, Trotta R, Butchar JP, Yu B, Benson DM Jr, Roda JM, Cheney C, Mo X, Lehman A, Jones J, Flynn J, Jarjoura D, Desjarlais JR, Tridandapani S, Caligiuri MA, Muthusamy N, Byrd JC. CD19 targeting of chronic lymphocytic leukemia with a novel Fc-domain-engineered monoclonal antibody. Blood. 2010 Feb 11;115(6):1204-13. doi: 10.1182/blood-2009-06-229039. Epub 2009 Dec 2. PMID: 19965644; PMCID: PMC2826232.