KC-5493

CHOK1-mouse-ALK7-Cell-Line

56254

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Background of CHOK1-mouse-ALK7-Cell-Line

The Activin A Receptor Type 1C (ACVR1C), also known as ALK-7, belongs to the transforming growth factor-beta (TGF-β) superfamily of serine/threonine kinase receptors. ACVR1C plays a significant role in various biological processes, including cell proliferation, differentiation, and apoptosis by mediating signals from activin A, a cytokine involved in numerous physiological functions. Research has indicated that ACVR1C is implicated in metabolic regulation, immune responses, and embryonic development. Dysregulation or mutations in this gene have been linked to several diseases, such as polycystic ovary syndrome (PCOS) and cancer.

Specifications

Catalog Number	KC-5493
Cell Line Name	CHOK1-mouse-ALK7-Cell-Line
NCBI/UniProt Accession Number	NM_001111030.1
Clone Number	3-9#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous mouse-ALK7 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% 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-mouse-ALK7-cell-line was generated using a lentiviral vector expressing the mouse-ALK7 sequence.

Characterization

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

Figure 2: Characterization of mouse-ALK7 in the CHOK1-mouse-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.