KC-1036

CTLL-2-Cell-Line-(Not-for-sale)

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Background of CTLL-2-Cell-Line-(Not-for-sale)

CTLL-2 is a clone of cytotoxic T cells derived from a C57BL/6 mouse. The cells are dependent upon Interleukin-2 (IL-2) for growth and can be used to assay for IL-2.

Specifications

Catalog Number	KC-1036
Cell Line Name	CTLL-2-Cell-Line-(Not-for-sale)
Host Cell Line	NA
Description	A subclone of T cells derived from a C57bl/6 mouse
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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+IL-2/RPMI1640+2 mM L-glutamine+ 1mM sodium pyruvate+ 10% fetal bovine serum+10% T-STIM with Con A
Selection Marker	NA
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 3-5 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Characterization

Cell Resuscitation

1. Prewarm culture medium (RPMI1640+IL-2/RPMI1640+2 mM L-glutamine+ 1mM sodium pyruvate+ 10% fetal bovine serum+10% T-STIM with Con A)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 1:4-1:5 every 2-3 days; seed out at about 3-5 × 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. Long term culture of tumour-specific cytotoxic T cells. Nature 268:154-156(1977)

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.