KC-4685

CHOK1-FZD10 Cell Line

47754

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Background of CHOK1-FZD10 Cell Line

FZD10 (Frizzled Class Receptor 10) is a Protein Coding gene, it is a member of the frizzled gene family. Members of this family encode 7-transmembrane domain proteins that are receptors for the Wingless type MMTV integration site family of signaling proteins. FZD proteins, a family of receptors comprised in the Wnt signaling pathway, play an important role in carcinogenesis of the gastroenteric tract. Diseases associated with FZD10 include Fallopian Tube Serous Adenocarcinoma and Colorectal Cancer.

Specifications

Catalog Number	KC-4685
Cell Line Name	CHOK1-FZD10 Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human FZD10 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

Cell Line Generation

CHOK1-FZD10 Cell Line was generated using a lentiviral vector expressing the human FZD10 sequence.

Characterization

Figure 1: Characterization of human FZD10 overexpression in the CHO-K1 FZD10 stable clone using QPCR.

Figure 2: Characterization of human FZD10 overexpression in the CHO-K1 FZD10 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

Ngernsombat, C., Suriya, U., Prattapong, P. et al. Repurposing FDA-approved drugs targeting FZD10 in nasopharyngeal carcinoma: insights from molecular dynamics simulations and experimental validation. Sci Rep 14, 31461 (2024).
Nagayama, S., Fukukawa, C., Katagiri, T. et al. Therapeutic potential of antibodies against FZD10, a cell-surface protein, for synovial sarcomas. Oncogene 24, 6201–6212 (2005).
Scavo MP, Depalo N, Rizzi F, Ingrosso C, Fanizza E, Chieti A, Messa C, Denora N, Laquintana V, Striccoli M, Curri ML, Giannelli G. FZD10 Carried by Exosomes Sustains Cancer Cell Proliferation. Cells. 2019 Jul 25;8(8):777. doi: 10.3390/cells8080777. PMID: 31349740; PMCID: PMC6721576.