KC-3870

CHOK1-cyno-CD22-Cell-Line

34936

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Background of CHOK1-cyno-CD22-Cell-Line

CD22 (CD22 Molecule) also known as SIGLEC2, it is a Protein Coding gene. Sialic acid-binding immunoglobulin-like lectin (Siglec) receptors are a family of 14 cell surface transmembrane proteins that bind specifically to sialic acid (Sia)-containing glycans, facilitating cell adhesion and/or cell signaling1. CD22 (Siglec 2) is a receptor predominantly restricted to B cells, it is a single-spanning membrane glycoprotein of 140 kDa on the surface of B cells. The cis/trans binding modes of CD22, together with a multi-Ig domain topology imply that the biological function of CD22 might be facilitated by significant conformational plasticity. Diseases associated with CD22 include Hematologic Cancer and Refractory Hairy Cell Leukemia.

Specifications

Catalog Number	KC-3870
Cell Line Name	CHOK1-cyno-CD22-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous cyno CD22 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

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

Characterization

Figure 1: Characterization of cyno CD22 overexpression in the CHO-K1 cyno CD22 stable clone using QPCR.

Figure 2: Characterization of cyno CD22 overexpression in the CHO-K1 cyno CD22 stable clone using PCR sequence.

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 1:4-1:8 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

Pluvinage JV, Haney MS, Smith BAH, Sun J, Iram T, Bonanno L, Li L, Lee DP, Morgens DW, Yang AC, Shuken SR, Gate D, Scott M, Khatri P, Luo J, Bertozzi CR, Bassik MC, Wyss-Coray T. CD22 blockade restores homeostatic microglial phagocytosis in ageing brains. Nature. 2019 Apr;568(7751):187-192. doi: 10.1038/s41586-019-1088-4. Epub 2019 Apr 3. PMID: 30944478; PMCID: PMC6574119.
Clark EA, Giltiay NV. CD22: A Regulator of Innate and Adaptive B Cell Responses and Autoimmunity. Front Immunol. 2018 Sep 28;9:2235. doi: 10.3389/fimmu.2018.02235. PMID: 30323814; PMCID: PMC6173129.
Ereño-Orbea, J., Sicard, T., Cui, H. et al. Molecular basis of human CD22 function and therapeutic targeting. Nat Commun 8, 764 (2017).