KC-3258

293T-mouse-TFRC Cell Line

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Background of 293T-mouse-TFRC Cell Line

TFRC, also known as CD71, is a transmembrane glycoprotein that encodes a cell surface receptor. TfR1 is required for iron import from transferrin into cells by endocytosis. TFRC is a potential new target in cases of human leukomia & lymphoma. TFRC has been shown to interact with GABARAP and HFE.

Specifications

Catalog Number	KC-3258
Cell Line Name	293T-mouse-TFRC Cell Line
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous mouse-TFRC 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 1:4-1:8 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
In Vivo Validation	NA

Cell Line Generation

293T-mouse-TFRC cell line was generated using lentivirus expressing mouse-TFRC sequence.

Characterization

Figure 1. Characterization of mouse-TFRC over-expression in the 293T-mouse-TFRC stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 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 1:4-1:8 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

Antineoplastic efficacy profiles of avapritinib and nintedanib in KIT D816V+ systemic mastocytosis: a preclinical study. Degenfeld-Schonburg, Gamperl, Stefanzl et al. Am J Cancer Res (2023) 13 (2), 355-378.
Aisen P (November 2004). "Transferrin receptor 1". The International Journal of Biochemistry & Cell Biology. 36 (11): 2137–43. doi:10.1016/j.biocel.2004.02.007. PMID 15313461.
Moos T (November 2002). "Brain iron homeostasis". Danish Medical Bulletin. 49 (4): 279–301. PMID 12553165
Speeckaert MM, Speeckaert R, Delanghe JR (December 2010). "Biological and clinical aspects of soluble transferrin receptor". Critical Reviews in Clinical Laboratory Sciences. 47 (5–6): 213–28. doi:10.3109/10408363.2010.550461. PMID 21391831. S2CID 25425279.