KC-6441

293T-DKK1-Cell-Line

63310

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Background of 293T-DKK1-Cell-Line

DKK1 encodes a member of the dickkopf protein family. Members of this family are secreted proteins characterized by two cysteine-rich domains that mediate protein-protein interactions. The encoded protein can bind the LRP6 co-receptor and inhibit β-catenin-dependent Wnt signaling pathways. This gene plays a role in embryonic development and may be important in adult bone formation. Elevated expression of this gene has been observed in various human cancers, and the protein may promote proliferation, invasion, and growth of cancer cell lines.

Specifications

Catalog Number	KC-6441
Cell Line Name	293T-DKK1-Cell-Line
NCBI/UniProt Accession Number	NM_012242.4
Clone Number	3#
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous DKK1 gene
Quantity	One vial 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 + 1μg/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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-DKK1-cell-line was generated using a lentiviral vector expressing the DKK1 sequence.

Characterization

Figure 1: Characterization of DKK1 overexpression in the 293T-DKK1 stable clone using qPCR.

Figure 2: Characterization of DKK1 in the 293T-DKK1 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage.

References

1.Ma L, Yang L, Wen J, Lu F, Yu J, Meng X. DKK1 expression associates with tertiary lymphoid structure maturity in non-small cell lung cancer. Apoptosis. 2026 Mar 29;31(4):116. doi: 10.1007/s10495-026-02332-z. PMID: 41904774.
2.Xu S, Liang L, Xu H, Wang Q, Wei K, Wang Z, Kong X. Decoding the DKK1/CKAP4 signaling Axis: A novel mechanism driving Neointimal hyperplasia in arteriovenous fistulas. Int Immunopharmacol. 2026 May 15;177:116520. doi: 10.1016/j.intimp.2026.116520. Epub 2026 Mar 21. PMID: 41865456.
3.Hie M, Iitsuka N, Otsuka T, Tsukamoto I. [Expression of Concern] Insulin‑dependent diabetes mellitus decreases osteoblastogenesis associated with the inhibition of Wnt signaling through increased expression of Sost and Dkk1 and inhibition of Akt activation. Int J Mol Med. 2026 May;57(5):128. doi: 10.3892/ijmm.2026.5799. Epub 2026 Mar 20. PMID: 41860049; PMCID: PMC13007994.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.