KC-3356

293T-CRTAM-Cell-Line

34346

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Background of 293T-CRTAM-Cell-Line

CRTAM, also known as CD355, is a class I MHC restricted T cell related molecule in the immunoglobulin superfamily, highly expressed in the cerebellum, especially in Purkinje neurons.CRTAM expression is restricted to activated class-I MHC-restricted T cells, including NKT and CD8 T cells. CRTAM represents one of the most highly expressed surface markers of activated human CD8 T cells and NKT cells, suggesting it may have diagnostic uses in various human viral and autoimmune diseases.

Specifications

Catalog Number	KC-3356
Cell Line Name	293T-CRTAM-Cell-Line
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous CRTAM gene in high level
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
In Vivo Validation	NA

Cell Line Generation

293T-CRTAM cell line was generated using a lentiviral vector expressing the CRTAM sequence.

Characterization

Figure 1: Characterization of CRTAM overexpression in the 293T-CRTAM stable clone using FACS.

Figure 2: Characterization of CRTAM in the 293T-CRTAM 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

Garay E, Patiño-López G, Islas S, Alarcón L, Canche-Pool E, Valle-Rios R, Medina-Contreras O, Granados G, Chávez-Munguía B, Juaristi E, Ortiz-Navarrete V, González-Mariscal L. CRTAM: A molecule involved in epithelial cell adhesion. J Cell Biochem. 2010 Sep 1;111(1):111-22. doi: 10.1002/jcb.22673. PMID: 20556794.
Patiño-Lopez G, Hevezi P, Lee J, Willhite D, Verge GM, Lechner SM, Ortiz-Navarrete V, Zlotnik A. Human class-I restricted T cell associated molecule is highly expressed in the cerebellum and is a marker for activated NKT and CD8+ T lymphocytes. J Neuroimmunol. 2006 Feb;171(1-2):145-55. doi: 10.1016/j.jneuroim.2005.09.017. Epub 2005 Nov 18. PMID: 16300832.

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.