KC-3354

CHOK1 mouse CD22 Middle Cell Line

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Background of CHOK1 mouse CD22 Middle Cell Line

CD22 (CD22 Molecule) is a Protein Coding gene,This gene involved in several processes, including negative regulation of calcium-mediated signaling; negative regulation of immunoglobulin production; and regulation of B cell proliferation. It is integral component of plasma membrane.This gene is expressed in several structures, including alimentary system; brain; hemolymphoid system gland; liver and biliary system; and reproductive system. Diseases associated with CD22 include Refractory Hairy Cell Leukemia and Refractory Hematologic Cancer.

Specifications

Catalog Number	KC-3354
Cell Line Name	CHOK1 mouse CD22 Middle Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous mouse CD22 gene in middle level
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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 mouse-CD22-middle Cell Line was generated using a lentiviral vector expressing the mouse-CD22 sequence.

Characterization

Figure 1: Characterization of mouse-CD22 overexpression in the CHOK1 mouse-CD22 stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/mL Puromycin)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 1:4-1:5 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

Dörner T, Shock A, Smith KG. CD22 and autoimmune disease. Int Rev Immunol. 2012 Oct;31(5):363-78. doi: 10.3109/08830185.2012.709890. PMID: 23083346.
Walker JA, Smith KG. CD22: an inhibitory enigma. Immunology. 2008 Mar;123(3):314-25. doi: 10.1111/j.1365-2567.2007.02752.x. Epub 2007 Dec 7. PMID: 18067554; PMCID: PMC2433339.
Cesano A, Gayko U. CD22 as a target of passive immunotherapy. Semin Oncol. 2003 Apr;30(2):253-7. doi: 10.1053/sonc.2003.50057. PMID: 12720147.