KC-5684

293T-FLT1-Cell-Line

57707

Home » 293T-FLT1-Cell-Line

Background of 293T-FLT1-Cell-Line

FLT1 encodes a member of the vascular endothelial growth factor receptor (VEGFR) family. VEGFR family members are receptor tyrosine kinases (RTKs) which contain an extracellular ligand-binding region with seven immunoglobulin (Ig)-like domains, a transmembrane segment, and a tyrosine kinase (TK) domain within the cytoplasmic domain. This protein binds to VEGFR-A, VEGFR-B and placental growth factor and plays an important role in angiogenesis and vasculogenesis. Expression of this receptor is found in vascular endothelial cells, placental trophoblast cells and peripheral blood monocytes.

Specifications

Catalog Number	KC-5684
Cell Line Name	293T-FLT1-Cell-Line
Clone Number	6#
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous FLT1 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

Cell Line Generation

293T-FLT1-cell-line was generated using a lentiviral vector expressing the FLT1 sequence.

Characterization

Figure 1: Characterization of FLT1 overexpression in the 293T-FLT1 stable clone using FACS.(Primary antibody: Icrucumab, Cat#KB-1752, Kyinno)

Figure 2: Characterization of FLT1 overexpression in the 293T stable clone using FACS.

Figure 3: Characterization of FLT1 in the 293T-FLT1 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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.Gill A, Kinghorn K, Bautch VL, Mac Gabhann F. Mechanistic computational modeling of sFLT1 secretion dynamics. PLoS Comput Biol. 2025 Aug 18;21(8):e1013324. doi: 10.1371/journal.pcbi.1013324. PMID: 40825055; PMCID: PMC12370208.
2.Chen H, Yang J, Li Z, Gao F. FLT1 as a Protective Factor in Ischemic Stroke: Insights from Real-World Pharmacovigilance and Genetic Evidence. Transl Stroke Res. 2025 Jul 18. doi: 10.1007/s12975-025-01369-7. Epub ahead of print. PMID: 40679760.
3.Wang L, Zhao B, Wang J, Zhang D, Ma R, Zhang T, Qi Y, Sheng Y, Hu B, Jin T. Epigollatecatechin gallate alleviates rheumatoid arthritis through PI3K-Akt pathway by inhibiting FLT1. Int Immunopharmacol. 2025 Jul 28;160:114958. doi: 10.1016/j.intimp.2025.114958. Epub 2025 May 30. PMID: 40449273.