KC-5112

CHOK1-cyno-ALK7-Cell-Line

62515

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Background of CHOK1-cyno-ALK7-Cell-Line

ACVR1C (Activin A Receptor Type 1C) also known as ALK7, ALK7 (Activin receptor-like kinase 7) is a member of the TGF-β receptor superfamily predominantly expressed by cells and tissues involved in endocrine functions, such as neurons of the hypothalamus and pituitary, pancreatic β-cells and adipocytes. The TGFß superfamily receptor ALK7 is a suppressor of tumorigenesis and metastasis, Activation in neoplastic cells of the ALK7 signaling pathway by its principal ligand activin B induces apoptosis. Diseases associated with ACVR1C include Extraocular Retinoblastoma and Acquired Hyperkeratosis.

Specifications

Catalog Number	KC-5112
Cell Line Name	CHOK1-cyno-ALK7-Cell-Line
NCBI/UniProt Accession Number	XM_005573224.4
Clone Number	8-2#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous cyno-ALK7 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% RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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-cyno-ALK7-cell-line was generated using a lentiviral vector expressing the cyno-ALK7 sequence.

Characterization

Figure 1: Characterization of cyno-ALK7 overexpression in the CHOK1-cyno-ALK7 stable clone using FACS.

Figure 2: Characterization of cyno-ALK7 in the CHOK1-cyno-ALK7 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 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 1:6-1:8 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.Ibáñez CF. Regulation of metabolic homeostasis by the TGF-β superfamily receptor ALK7. FEBS J. 2022 Oct;289(19):5776-5797. doi: 10.1111/febs.16090. Epub 2021 Jul 11. PMID: 34173336.
2.Gong FH, Cheng WL, Zhang Q, Chen XL, Cao JL, Yang T, Song WH, Zhao F. ALK7 Promotes Vascular Smooth Muscle Cells Phenotypic Modulation by Negative Regulating PPARγ Expression. J Cardiovasc Pharmacol. 2020 Aug;76(2):237-245. doi: 10.1097/FJC.0000000000000857. PMID: 32467530.
3.Michael IP, Saghafinia S, Tichet M, Zangger N, Marinoni I, Perren A, Hanahan D. ALK7 Signaling Manifests a Homeostatic Tissue Barrier That Is Abrogated during Tumorigenesis and Metastasis. Dev Cell. 2019 May 6;49(3):409-424.e6. doi: 10.1016/j.devcel.2019.04.015. PMID: 31063757.
4.Goebel EJ, Ongaro L, Kappes EC, Vestal K, Belcheva E, Castonguay R, Kumar R, Bernard DJ, Thompson TB. The orphan ligand, activin C, signals through activin receptor-like kinase 7. Elife. 2022 Jun 23;11:e78197. doi: 10.7554/eLife.78197. PMID: 35736809; PMCID: PMC9224996.

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.