KC-2426

293T-CD36-Cell-Line

24036

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Background of 293T-CD36-Cell-Line

CD36 belongs to the family of group B scavenger receptors, a highly glycosylated dual transmembrane protein, also known as fatty acid transporter enzyme, platelet membrane glycoprotein IV, serum granule protein, and class B type 2 scavenger receptor, is a multifunctional scavenger receptor with a variety of ligand recognition sites, dominates the fatty acid transmembrane transport process, and plays an important role in lipid metabolism.

Specifications

Catalog Number	KC-2426
Cell Line Name	293T-CD36-Cell-Line
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous human CD36 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-CD36-cell-line was generated using a lentiviral vector expressing the CD36 sequence.

Characterization

Figure 1: Characterization of CD36 overexpression in the 293T-CD36 stable clone using FACS.

Figure 2: Characterization of CD36 overexpression in the 293T stable clone using FACS.

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.Glatz JFC, Nabben M, Luiken JJFP. CD36(SR- B2) as master regulator of cellular fatty acid homeostasis. Current Opinion in Lipid ology, 33(2):103- 111 (2022).
2.Feng WW, Wilkins O, et al. CD36-Mediated Metabolic Rewiring of Breast Cancer Cells Promotes Resistance to HER2-Targeted Therapies. Cell Reports, 29(11):3405-3420 (2019).
3.Wang H, Franco F, et al. CD36-mediated metabolic adaptation supports regulatory T cell survival and function in tumors. Nature Immunology, 21(3):298-308 (2020).