KC-3181

293T-TFRC-Cell-Line

Stable HEK293T cell line expressing exogenous human TFRC gene

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Background of 293T-TFRC-Cell-Line

TFRC, also known as CD71, is a transmembrane glycoprotein that encodes a cell surface receptor. TFRC 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-3181
Cell Line Name	293T-TFRC-Cell-Line
Host Cell Line	293T
Description	Stable HEK293T cell line expressing exogenous human 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 human TFRC cell line was generated using a lentiviral vector expressing the human TFRC sequence.

Characterization

Figure1: Characterization of human TFRC overexpression in the 293T human TFRC stable clone using FACS.

Figure2: Characterization of human TFRC expression in 293T cells using FACS.

Figure3: Characterization of human TFRC expression in the 293T human TFRC stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 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–
doi:10.3109/10408363.2010.550461. PMID 21391831. S2CID 25425279.