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KC-1934

CHOK1-CD94-NKG2A-Cell-Line

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Home » 细胞系 » CHOK1-CD94-NKG2A-Cell-Line

Background of CHOK1-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.NKG2-A 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. Ba/F3 cell, a murine interleukin-3 dependent pro-B cell line, is a popular system for exploring both kinases and their inhibitors, because some protein kinases can render the Ba/F3 cells to be depended on the activation of the kinases instead of IL-3 supplement, while their inhibitors can antagonize the kinase-dependent growth effects.

Specifications

Catalog NumberKC-1934
Cell Line NameCHOK1-CD94-NKG2A-Cell-Line
Host Cell LineChinese hamster ovary CHO-K1 cell line
DescriptionStable CHOK1 cell line 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-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

CHOK1 human CD94 NKG2A cell line was generated using a lentiviral vector expressing the human CD94 and NKG2A sequence.

Characterization

Cell Resuscitation

  1. Prewarm culture medium (RPMI1640 + 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% RPMI-1640 + 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. Borrego F, Masilamani M, Marusina AI, Tang X, Coligan JE (2006), The CD94/NKG2 family of receptors: from molecules and cells to clinical relevance. Immunol Res. 35(3):263-78
  2. CD94 surface density identifies a functional intermediary between the CD56bright and CD56dim human NKcell subsets. Jianhua Yu, Hsiaoyin C. Mao, Min Wei, Tiffany Hughes, Jianying Zhang, Il-kyoo Park, Shujun Liu, Susan McClory, Guido Marcucci, Rossana Trotta, Michael A. CaligiuriBlood (2010) 115 (2): 274ÿ281.
  3. Borrego F, Kabat J, Kim DK, Lieto L, Maasho K, Pena J et al. Structure and function of major histocompatibility complex (MHC) class I specific receptors expressed on human natural killer (NK) cells. Mol Immunol 2002; 38: 637ÿ660.

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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