KC-2293-DW

CT26-GDF15 Cell Line

61075

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Background of CT26-GDF15 Cell Line

GDF15, also named MIC-1, is a member of TGF-beta superfamily proteins, and plays an important role in tumorigenesis and metastasis. the expression of GDF-15 is dramatically increased in many types of cancers, such as colorectal, breast, and prostate. GDF-15 also mediates cancer-induced anorexia and weight loss through the modulation of neuronal pathways important in the regulation of appetite and energy homeostasis.

Specifications

Catalog Number	KC-2293-DW
Cell Line Name	CT26-GDF15 Cell Line
NCBI/UniProt Accession Number	NM_004864.4
Clone Number	3#
Host Cell Line	CT26
Description	Stable CT26-GDF15 cell line expressing exogenous GDF15 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 1:10 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 20 hours
Mycoplasma Status	Negative

Cell Line Generation

CT26-GDF15 cell line was generated using lentivirus expressing GDF15 sequence.

Characterization

Figure 1. Characterization of GDF15 over-expression in the CT26-GDF15 stable clone using Elisa.

Figure 2:Validation of in vivo tumorigenicity of CT26 cells stably expressing GDF15 via subcutaneous implantation in BALB/c mice, with tumor growth monitored by measuring volume (V=0.5×length×width²) and body weight.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS+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 1:10 every 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

GDF15, an update of the physiological and pathological roles it plays: a review.Artin Assadi , Azadeh Zahabi&Robert A Hart .Affiliations expand PMID: 32936319
Secreted and Cell Surface Genes Expressed in Benign and Malignant Colorectal Tumors. Advances in Brief DOI: Published October 2001 doi:10.1006/geno.1998.5281. PMID 9615235.
The Transforming Growth Factor-β Superfamily Member Growth-Differentiation Factor-15 Protects the Heart From Ischemia/Reperfusion Injury.Circulation Research. 2006;98:351-360.Originally published5 Jan 2006.