KC-2367

CHOK1-cyno-CD7-Cell-Line

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Home » 细胞系 » CHOK1-cyno-CD7-Cell-Line

Background of CHOK1-cyno-CD7-Cell-Line

CD7 antigen is a member of the immunoglobulin superfamily, which is a member of the immunoglobulin superfamily. Expressed on the surface of all thymocytes, most mature T cells, and natural killer cells. It plays an important role in T cell interactions and T cell/B cell interactions during early lymphatic development.

Specifications

Catalog NumberKC-2367
Cell Line NameCHOK1-cyno-CD7-Cell-Line
Host Cell LineCHOK1
DescriptionStable CHOK1 clone expressing exogenous cyno-CD7 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% RPMI 1640 + 20% FBS + 10% DMSO
Propagation MediumRPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial
SubcultureSplit saturated culture 1:6-1:8 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

CHOK1-cyno-CD7-cell-line was generated using a lentiviral vector expressing the cyno-CD7 sequence.

Characterization

Figure 1: Characterization of cyno-CD7 overexpression in the CHOK1-cyno-CD7 stable clone using qPCR.

Cell Resuscitation

  1. Prewarm culture medium (RPMI 1640 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 1:6-1:8 every 2-3 days; seed out at about 1-3 × 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.Haftcheshmeh SM, Tajbakhsh A, Kazemi M, Esmaeili SA, Mardani F, Fazeli M, Sahebkar A. The clinical importance of CD4+ CD7- in human diseases. J Cell Physiol. 2019 Feb;234(2):1179-1189. doi: 10.1002/jcp.27099. Epub 2018 Aug 1. PMID: 30067877. 2.Stillwell R, Bierer BE. T cell signal transduction and the role of CD7 in costimulation. Immunol Res. 2001;24(1):31-52. doi: 10.1385/ir:24:1:31. PMID: 11485208.

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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