KC-6293

HT1080-GDF15-KO cell line

63956

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Background of HT1080-GDF15-KO cell line

Growth differentiation factor 15 (GDF15), also known as macrophage inhibitory cytokine-1 (MIC-1), is located on the forward strand of the short arm of chromosome 19 (19p13.11). It is a stress response cytokine expressed in a variety of tissue and secreted into circulation in response to many stimuli as part of a wide variety of disease processes, including cancer and obesity.

Specifications

Catalog Number	KC-6293
Cell Line Name	HT1080-GDF15-KO cell line
Clone Number	2A4
Host Cell Line	HT1080
Description	Stable HT1080 cell line with human GDF15 gene knockout
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	DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS
Selection Marker	NA
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split saturated culture 1:4-1:5 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

Cell Line Generation

HT1080-GDF15-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of HT1080-GDF15-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of HT1080-GDF15-KO cell line stable clone using RT-PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM+10% FBS) 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 1:4-1:5 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

Borner T, Shaulson ED, Ghidewon MY, Barnett AB, Horn CC, Doyle RP, Grill HJ, Hayes MR, De Jonghe BC. GDF15 Induces Anorexia through Nausea and Emesis. Cell Metab. 2020 Feb 4;31(2):351-362.e5. doi: 10.1016/j.cmet.2019.12.004. Epub 2020 Jan 9. PMID: 31928886; PMCID: PMC7161938.
Lockhart SM, Saudek V, O'Rahilly S. GDF15: A Hormone Conveying Somatic Distress to the Brain. Endocr Rev. 2020 Aug 1;41(4):bnaa007. doi: 10.1210/endrev/bnaa007. PMID: 32310257; PMCID: PMC7299427.

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.