KC-1036

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

21423

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

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	Two vials 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)