KC-3099

CHOK1-TMEFF2-High Cell Line

34126

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Background of CHOK1-TMEFF2-High Cell Line

TMEFF2 (Transmembrane protein with EGF-like and two follistatin-like domains 2), also known as TENB2 or TPEF, is a type-I transmembrane proteoglycan primarily expressed in the prostate and central nervous system. It plays a complex dual role in cell signaling, acting as a neuroprotective survival factor in the brain while modulating the AKT and MAPK pathways in malignancies. In prostate cancer, TMEFF2 is significantly overexpressed and androgen-regulated, making it a critical diagnostic biomarker and therapeutic target. Current drug development focuses on Antibody-Drug Conjugates (ADCs) and bispecific T-cell engagers (TCEs); for instance, recent clinical trials for candidates like JNJ-70218902 (a T-cell redirecting antibody) have explored its potential in treating metastatic castration-resistant prostate cancer (mCRPC).

Specifications

Catalog Number	KC-3099
Cell Line Name	CHOK1-TMEFF2-High Cell Line
NCBI/UniProt Accession Number	NM_016192.4
Clone Number	3#
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human TMEFF2 gene in high level
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% 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 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-TMEFF2-High Cell Line was generated using a lentiviral vector expressing the human TMEFF2 sequence.

Characterization

Figure 1: Characterization of human TMEFF2 overexpression in the CHOK1-TMEFF2 stable high level clone using FACS.

Figure2: Characterization of endogenous human TMEFF2 expression in CHOK1 using FACS.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 + 10% FBS + 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 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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, Motasim et al. “TMEFF2: A Transmembrane Proteoglycan with Multifaceted Actions in Cancer and Disease.” Cancers vol. 12,12 3862. 21 Dec. 2020, doi:10.3390/cancers12123862
2. Gao, Lingling et al. “TMEFF2 is a novel prognosis signature and target for endometrial carcinoma.” Life sciences vol. 243 (2020): 116910. doi:10.1016/j.lfs.2019.116910
3. Xie, Sidi et al. “TMEFF2 promoter hypermethylation is an unfavorable prognostic marker in gliomas.” Cancer cell international vol. 21,1 148. 4 Mar. 2021, doi:10.1186/s12935-021-01818-x
4. Calvo, Emiliano et al. “A first-in-human study of JNJ-70218902, a bispecific T-cell-redirecting antibody against TMEFF2 in metastatic castration-resistant prostate cancer.” The oncologist vol. 30,1 (2025): oyae313. doi:10.1093/oncolo/oyae313

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.