KC-3239

CHOK1-cyno-TMEFF2 Cell Line

34248

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Background of CHOK1-cyno-TMEFF2 Cell Line

TMEFF2 encodes a member of the tomoregulin family of transmembrane proteins. This protein has been shown to function as both an oncogene and a tumor suppressor depending on the cellular context and may regulate prostate cancer cell invasion. Multiple soluble forms of this protein have been identified that arise from both an alternative splice variant and ectodomain shedding. Additionally, this gene has been found to be hypermethylated in multiple cancer types. Alternative splicing results in multiple transcript variants.

Specifications

Catalog Number	KC-3239
Cell Line Name	CHOK1-cyno-TMEFF2 Cell Line
NCBI/UniProt Accession Number	XM_005573793.3
Clone Number	27#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous cyno-TMEFF2 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 1:3~1:6 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
In Vivo Validation	NA

Cell Line Generation

CHOK1-cyno-TMEFF2 cell Line was generated using retrovirus vector expressing cyno-TMEFF2 sequence.

Characterization

Figure 1: Characterization of CHOK1-cyno-TMEFF2 Cell Line stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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 1:3~1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI-1640 + 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.Masood M, Grimm S, El-Bahrawy M, Yagüe E. TMEFF2: A Transmembrane Proteoglycan with Multifaceted Actions in Cancer and Disease. Cancers (Basel). 2020 Dec 21;12(12):3862. doi: 10.3390/cancers12123862. PMID: 33371267; PMCID: PMC7766544.
2.Xie S, Zhang Y, Peng T, Guo J, Cao Y, Guo J, Shi X, Li Y, Liu Y, Qi S, Wang H. TMEFF2 promoter hypermethylation is an unfavorable prognostic marker in gliomas. Cancer Cell Int. 2021 Mar 4;21(1):148. doi: 10.1186/s12935-021-01818-x. PMID: 33663520; PMCID: PMC7931334.
3.Gao L, Nie X, Zheng M, Li X, Guo Q, Liu J, Liu Q, Hao Y, Lin B. TMEFF2 is a novel prognosis signature and target for endometrial carcinoma. Life Sci. 2020 Feb 15;243:116910. doi: 10.1016/j.lfs.2019.116910. Epub 2019 Oct 11. PMID: 31610211.