KC-4597

NCI-H1436-Cell-Line-(Not-for-sale)

52725

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Background of NCI-H1436-Cell-Line-(Not-for-sale)

NCI-H1436 [H1436] is a cell line that was isolated from the lungs of a 39-year-old, White male with stage E small cell lung cancer. This product can be used for cancer research.

Specifications

Catalog Number	KC-4597
Cell Line Name	NCI-H1436-Cell-Line-(Not-for-sale)
Host Cell Line	NA
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	3D cell culture; Cancer research
Freezing Medium	70% DMEM/F12K(1:1)+20% FBS+10% DMSO
Propagation Medium	DMEM/F12K(1:1)+ ITS+10 nM Hydrocortisone +10 nM beta-estradiol +5% FBS(Gibco)
Selection Marker	NA
Morphology	Growing in clumps in suspension
Subculture	Split saturated culture 1:2-1:3 every 4-5 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 100 hours
Mycoplasma Status	Negative

Cell Line Generation

Characterization

Cell Resuscitation

1. Prewarm culture medium (DMEM/F12K(1:1)+ ITS+10 nM Hydrocortisone +10 nM beta-estradiol +5% FBS(Gibco))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 1:2-1:3 every 4-5 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM/F12K(1:1)+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. NCI-Navy Medical Oncology Branch cell line data base. J. Cell. Biochem. Suppl. 24:32-91(1996)
2. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 483:603-607(2012)