KC-2411

HEK293T-cyno-TF-Cell-Line

24004

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Background of HEK293T-cyno-TF-Cell-Line

TF, also known as CD142, is a type I transmembrane glycoprotein, which is expressed on the surface of a variety of cells that are physically separated from the circulating blood which include smooth muscle cells, fibroblasts, keratinocytes, glomerular epithelial cells (cytoplasmic inclusions), astrocytes, myocardium, liver stromal cells, pancreas cells, and is also expressed on activated monocytes and stimulated endothelial cells. TF is a high-affinity receptor for coagulation factor VII and initiates the extrinsic pathway of blood coagulation. TF also plays an important role in a variety of diseases such as sepsis, atherosclerosis, and cancer.

Specifications

Catalog Number	KC-2411
Cell Line Name	HEK293T-cyno-TF-Cell-Line
Host Cell Line	Human HEK293T cell line
Description	Stable HEK293T cell line expressing exogenous cyno TF gene
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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 every 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 cyno TF cell line was generated using a lentiviral vector expressing the cyno TF sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 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 1:3-1:6 every 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

"Entrez Gene: F3 coagulation factor III (thromboplastin, tissue factor)".
Muller YA, Ultsch MH, de Vos AM (February 1996). "The crystal structure of the extracellular domain of human tissue factor refined to 1.7 A resolution". Journal of Molecular Biology. 256 (1): 144 ÿ 59. doi:10.1006/jmbi.1996.0073. PMID 8609606.
Zhang E, St Charles R, Tulinsky A (February 1999). "Structure of extracellular tissue factor complexed with factor VIIa inhibited with a BPTI mutant". Journal of Molecular Biology. 285 (5): 2089 ÿ 104. doi:10.1006/jmbi.1998.2452. PMID 9925787.
Tissue factor in hematological malignancies. Versteeg, H.H. et al. (2003) Carcinogenesis 24:1009.