KC-5060

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

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

ACVR2A (Activin Receptor Type-2A) is a gene that encodes a key transmembrane receptor for the TGF-beta (Transforming Growth Factor-beta) superfamily.Enables several functions, including PDZ domain binding activity; activin binding activity; and transmembrane receptor protein serine/threonine kinase activity. Involved in BMP signaling pathway. Acts upstream of or within several processes, including Sertoli cell proliferation; copulation; and regionalization. Located in cell surface. Is active in plasma membrane. Is expressed in several structures, including central nervous system; early conceptus; gonad; gut; and sensory organ. Used to study Weissenbacher-Zweymuller syndrome. Human ortholog(s) of this gene implicated in Lynch syndrome and colon cancer. Orthologous to human ACVR2A (activin A receptor type 2A).

Specifications

Catalog Number	KC-5060
Cell Line Name	293T-ACVR2A-ACVR2B-KO-mouse-ACVR2A- Cell Line
NCBI/UniProt Accession Number	NM_007396.4
Clone Number	2#
Host Cell Line	293T
Description	Stable 293T-ACVR2A-ACVR2B-KO cell line expressing exogenous mouse-ACVR2A 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-mouse-ACVR2A cell line was generated using lentivirus expressing mouse-ACVR2A sequence.

Characterization

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

Figure 2. Characterization of 293T-ACVR2A-ACVR2B-KO-mouse-ACVR2A 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

Ihn HJ, Kim DH, Oh SS, Moon C, Chung JW, Song H, Kim KD. Identification of Acvr2a as a Th17 cell-specific gene induced during Th17 differentiation. Biosci Biotechnol Biochem. 2011;75(11):2138-41. doi: 10.1271/bbb.110436. Epub 2011 Nov 7. PMID: 22056434.

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.