KC-1803

MC38-CD24-Cell-Line

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Background of MC38-CD24-Cell-Line

CD24 encodes a salivary glycoprotein that is expressed on mature granulocytes and B cells, and regulates the growth and differentiation signals of these cells. It is also expressed in various malignant tumor cells, especially in ovarian cancer cells and ovarian cancer stem cells. CD24 expression is associated with increased metastatic potential and poor prognosis in malignant tumors. CD24 on the surface of tumor cells can interact with Siglec-10 on the surface of immune cells, mediating immune escape of tumor cells.

Specifications

Catalog Number	KC-1803
Cell Line Name	MC38-CD24-Cell-Line
Host Cell Line	MC38
Description	Stable MC38 clone expressing exogenous CD24 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 5μg/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 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

MC38-CD24-cell-line was generated using a lentiviral vector expressing the CD24 sequence.

Characterization

Figure 1: Characterization of CD24 overexpression in the MC38-CD24 stable clone using FACS.

Cell Resuscitation

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

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Gu Y, Zhou G, Tang X, Shen F, Ding J, Hua K. The biological roles of CD24 in ovarian cancer: old story, but new tales. Front Immunol. 2023 Jun 9;14:1183285. doi: 10.3389/fimmu.2023.1183285. PMID: 37359556; PMCID: PMC10288981.
Li X, Tian W, Jiang Z, Song Y, Leng X, Yu J. Targeting CD24/Siglec-10 signal pathway for cancer immunotherapy: recent advances and future directions. Cancer Immunol Immunother. 2024 Jan 27;73(2):31. doi: 10.1007/s00262-023-03606-0. PMID: 38279998; PMCID: PMC10821995.