KC-4577

293T-Norrin Cell Line

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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	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-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.

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.