KC-1482

MC38-Siglec15-Cell-Line

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Background of MC38-Siglec15-Cell-Line

Siglec-15 is a transmembrane glycoprotein, which belongs to the immunoglobulin superfamily and SIGLEC (sialic acid binding Ig-like lectin) family, and is found primarily on the surface of immune cells. Siglec-15 is that recognizes sialylated glycans and regulates osteoclast differentiation. Siglec-15 is a potential therapeutic target for osteoporosis and plays a conserved regulatory role in the immune system of vertebrates.

Specifications

Catalog Number	KC-1482
Cell Line Name	MC38-Siglec15-Cell-Line
Host Cell Line	Mouse MC38 cell line
Description	MC38 cell line stable expressing exogenous human siglec15 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+5µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:2-1:5 every 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 48 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

MC38 human siglec15 cell line was generated using lentiviral vector expressing human siglec15 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 5µg/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 1:2-1:5 every 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

Wang, Jun, Jingwei Sun, Linda N Liu, Dallas B Flies, Xinxin Nie, Maria Toki, Jianping Zhang, et al. 2019. ¡ùSiglec15 as an Immune Suppressor and Potential Target for Normalization Cancer Immunotherapy.¡ì Nature Medicine, March. Springer US, 1Ã¿32. doi:10.1038/s41591-019-0374-x.