kc-4190

293T-VPAC2 Cell Line

42619

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Background of 293T-VPAC2 Cell Line

VIPR2 (Vasoactive Intestinal Peptide Receptor 2) also known as VPAC2, it is a Protein Coding gene. Vasoactive intestinal peptide (VIP) receptors are a group of Gs-protein-coupled receptors, it is a small neuropeptide composing of 28 amino acids, can stimulate glucose-dependent insulin secretion, particularly by binding to VPAC2 receptors. The vasoactive intestinal peptide receptor 2 (VPAC2) is widely distributed throughout the body and is also overexpressed in a variety of human neoplastic tissues. VPAC2 has diverse biological actions including promotion of neuronal survival, regulation of glycogen metabolism, and inhibition of IL-2-mediated T-cell proliferation.

Specifications

Catalog Number	kc-4190
Cell Line Name	293T-VPAC2 Cell Line
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous human VPAC2 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
In Vivo Validation	NA

Cell Line Generation

Characterization

Figure 1: Characterization of VPAC2 overexpression in the 293T VPAC2 stable clone using QPCR.

Figure 2: Characterization of VPAC2 in the 293T VPAC2 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

Zhao X, Deng L, Ren L, Yang H, Wang B, Zhu X, Zhang X, Guo C, Zhang Y, Liu Y. VPAC2 receptor mediates VIP-potentiated insulin secretion via ion channels in rat pancreatic β cells. Exp Cell Res. 2023 Feb 15;423(2):113471. doi: 10.1016/j.yexcr.2023.113471. Epub 2023 Jan 13. PMID: 36642263.
Schulz S, Mann A, Novakhov B, Piggins HD, Lupp A. VPAC2 receptor expression in human normal and neoplastic tissues: evaluation of the novel MAB SP235. Endocr Connect. 2015 Mar;4(1):18-26. doi: 10.1530/EC-14-0051. Epub 2014 Dec 11. PMID: 25504760; PMCID: PMC4285768.
Hou X, Yang D, Yang G, Li M, Zhang J, Zhang J, Zhang Y, Liu Y. Therapeutic potential of vasoactive intestinal peptide and its receptor VPAC2 in type 2 diabetes. Front Endocrinol (Lausanne). 2022 Sep 20;13:984198. doi: 10.3389/fendo.2022.984198. PMID: 36204104; PMCID: PMC9531956.

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.