KC-5061

293T-ACVR2A-ACVR2B-KO-rat-ACVR2B- Cell Line

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Background of 293T-ACVR2A-ACVR2B-KO-rat-ACVR2B- Cell Line

ACVR2B (Activin Receptor Type-2B) is a gene that encodes a transmembrane receptor protein. It is part of the TGF-beta (Transforming Growth Factor-beta) superfamily signaling pathway. This receptor primarily binds to activins and other related ligands, such as myostatin, which are key regulators of growth and tissue homeostasis.

Specifications

Catalog Number	KC-5061
Cell Line Name	293T-ACVR2A-ACVR2B-KO-rat-ACVR2B- Cell Line
NCBI/UniProt Accession Number	NM_031554
Clone Number	1#
Host Cell Line	293T
Description	Stable 293T-ACVR2A-ACVR2B-KO cell line expressing exogenous rat-ACVR2B 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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS+1μg/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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-ACVR2A-ACVR2B-KO-rat-ACVR2B cell line was generated using lentivirus expressing rat-ACVR2B sequence.

Characterization

Figure 1. Characterization of rat-ACVR2B over-expression in the 293T-ACVR2A-ACVR2B-KO-rat-ACVR2B stable clone using FACS.

Figure 2. Characterization of 293T-ACVR2A-ACVR2B-KO-rat-ACVR2B cell line stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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% DMEM + 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

Feng B, Meng L, Luan L, Fang Z, Zhao P, Zhao G. Upregulation of Extracellular Vesicles-Encapsulated miR-132 Released From Mesenchymal Stem Cells Attenuates Ischemic Neuronal Injury by Inhibiting Smad2/c-jun Pathway via Acvr2b Suppression. Front Cell Dev Biol. 2021 Mar 8;8:568304. doi: 10.3389/fcell.2020.568304. PMID: 33763412; PMCID: PMC7982537.

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.