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KC-1937

CT26.WT-human-CD94-NKG2A Cell Line

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Datasheet
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Home » CT26.WT-human-CD94-NKG2A Cell Line

Background of CT26.WT-human-CD94-NKG2A Cell Line

CD94 & NKG2A, also known as KLRD1 & CD159A. CD94 is a type II integral membrane protein belonging to lectin superfamily, which is plays a role as a receptor for the recognition of MHC class I Human Leukocyte Antigen (HLA)-E molecules by NK cells and some cytotoxic T-cells. NKG2A contains C-type lectin domain and belongs to the killer cell lectin-like receptor (KLR) family. KLR family is a group of transmembrane proteins preferentially expressed in natural killer (NK) cells. CD94 can form disulfide-bonded heterodimer with NKG2A on the surface of NK cells. The CD94/NKG2A complex interacts with HLA-E on target cells and inhibit the cytotoxic activity of NK cells to prevent cell lysis.

Specifications

Catalog NumberKC-1937
Cell Line NameCT26.WT-human-CD94-NKG2A Cell Line
Host Cell LineCT26.WT
DescriptionCT26.WT cell line stable expressing exogenous human CD94 and NKG2A 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
SubcultureSplit saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 105 cells/ml
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative
In Vivo ValidationNA

Cell Line Generation

CT26.WT human CD94 NKG2A Cell Line was generated using a lentiviral vector expressing the CD94 and NKG2A sequence.

Characterization

Figure 1: Characterization of CD94 and NKG2A overexpression in the CT26.WT human CD94 NKG2A stable clone using FACS.

Cell Resuscitation

  1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 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 1-3 × 105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (70% RPMI1640 + 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. Turnpenny, P. D. et al. A gene for autosomal recessive spondylocostal dysostosis maps to 19q13.1-q13.3. The American Journal of Human Genetics 65, 175–182 (1999).
  2. Chapman, G., Sparrow, D. B., Kremmer, E. & Dunwoodie, S. L. Notch inhibition by the ligand Delta-Like 3 defines the mechanism of abnormal vertebral segmentation in spondylocostal dysostosis. Human Molecular Genetics 20, 905–916 (2010).
  3. Saunders, L. R. A DLL3-targeted antibody-drug conjugate eradicates high-grade pulmonary neuroendocrine tumor-initiating cells in vivo. Science Translational Medicine 7, 302ra136–302ra136 (2015).

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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