KC-4577

293T-Norrin Cell Line

47337

Home » 293T-Norrin Cell Line

Background of 293T-Norrin Cell Line

NDP (Norrin Cystine Knot Growth Factor NDP) also known as Norrin, it is a Protein Coding gene. Norrin is a secreted signaling molecule with structural and functional characteristics of an autocrine and/or paracrine acting growth factor. Norrin has pronounced neuroprotective properties on retinal ganglion cells (RGC) with the distinct potential to decrease the damaging effects of excitotoxic NMDA-induced RGC injury. Norrin mediates enhanced tumor growth of glioblastomas by activating the Notch pathway. Norrin inhibited cell growth via β-catenin signaling in GSCs that had low expression levels of the transcription factor ASCL1. Diseases associated with NDP include Norrie Disease and Exudative Vitreoretinopathy 2, X-Linked.

Specifications

Catalog Number	KC-4577
Cell Line Name	293T-Norrin Cell Line
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous human Norrin 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% 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-Norrin cell line was generated using a lentiviral vector expressing the human Norrin sequence.

Characterization

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

Figure 2: Characterization of human Norrin in the 293T Norrin 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

Ohlmann A, Tamm ER. Norrin: molecular and functional properties of an angiogenic and neuroprotective growth factor. Prog Retin Eye Res. 2012 May;31(3):243-57. doi: 10.1016/j.preteyeres.2012.02.002. Epub 2012 Feb 21. PMID: 22387751.
Díaz-Coránguez M, Lin CM, Liebner S, Antonetti DA. Norrin restores blood-retinal barrier properties after vascular endothelial growth factor-induced permeability. J Biol Chem. 2020 Apr 3;295(14):4647-4660. doi: 10.1074/jbc.RA119.011273. Epub 2020 Feb 21. PMID: 32086377; PMCID: PMC7135996.
Kassumeh S, Priglinger SG, Ohlmann A. Norrin mediates opposing effects on tumor progression of glioblastoma stem cells. J Clin Invest. 2020 Jun 1;130(6):2814-2815. doi: 10.1172/JCI137254. PMID: 32391807; PMCID: PMC7259986.