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KC-1189

CHOK1-cyno-TNFRSF1B-Cell-Line

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Datasheet
21675
Home » CHOK1-cyno-TNFRSF1B-Cell-Line

Background of CHOK1-cyno-TNFRSF1B-Cell-Line

Tumor necrosis factor (TNF) receptor 2 (TNFR2) is one of two receptors of the cytokines, TNF and lymphotoxin-the protective and anti-inflammatory effects of TNFR2 may explain why TNF inhibitors failed to be effective in diseases such as heart failure or multiple sclerosis, where TNF has been strongly implicated as a driving force. TNFR2 recruits the adapter protein TNF receptor-associated factor 2 (TRAF2) with the help short peptide motif and activates the alternative NFκB pathway.

Specifications

Catalog NumberKC-1189
Cell Line NameCHOK1-cyno-TNFRSF1B-Cell-Line
Host Cell LineChinese hamster ovary CHO-K1 cell line
DescriptionCHO-K1 cell line stable expressing exogenous cynomolgus TNFRSF1B gene
QuantityOne vial of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% RPMI1640+20% FBS+10% DMSO
Propagation MediumRPMI1640+10%FBS+10µg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial-like
SubcultureSplit saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 24 hours
Mycoplasma StatusNegative
In Vivo ValidationNA

Cell Line Generation

CHO-K1 Cyno TNFRSF1B cell line was generated using lentiviral vector expressing cynomolgus TNFRSF1B sequence

Characterization

Cell Resuscitation

  1. Prewarm culture medium (F12K + 10% FBS + 10µg/mL Puromycin)in a 37°C water bath.
  2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
  3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
  4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
  5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
  6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
  7. Incubate the flask at 37°C, 5% CO2 incubator.
  8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (70% F12K + 20% FBS + 10% DMSO) fresh immediately before use.
  2. Keep the freezing medium on ice and label cryovials.
  3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
  4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
  5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
  6. Aliquot 1 mL of the cell suspension into each cryovial.
  7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
  8. Transfer vials to liquid nitrogen for long-term storage

References

  1. WICOVSKY A, HENKLER F, SALZMANN S, et al. Tumor necrosis factor receptor-associated factor-1 enhances proinflammatory TNF receptor-2 signaling and modifies TNFR1-TNFR2 cooperation.[J]. Oncogene,2009,28(15):1769-1781.
  2. Pegoretti V, Baron W, Laman JD, et al. Selective modulation of TNF-TNFRs signaling: insights for multiple sclerosis treatment.Front Immunol.2018;9:925.

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.

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