KC-2475
293T-mouse-CD36-Cell-Line
Background of 293T-mouse-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-2475 |
|---|---|
| Cell Line Name | 293T-mouse-CD36-Cell-Line |
| Host Cell Line | 293T |
| Description | Stable 293T clone expressing exogenous mouse CD36 |
| 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-CD36-cell-line was generated using a lentiviral vector expressing the mouse-CD36 sequence.
Characterization
Figure 1: Characterization of mouse-CD36 overexpression in the 293T-mouse-CD36 stable clone using FACS.
Figure 2: Characterization of mouse-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.
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.
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.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).
2.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).
3.Tanase C, Enciu AM, et al. Fatty Acids, CD36, Thrombospondin-1, and CD47 in Glioblastoma: Together and/or Separately? International Journal of Molecular Sciences, 23(2):604 (2022).
2.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).
3.Tanase C, Enciu AM, et al. Fatty Acids, CD36, Thrombospondin-1, and CD47 in Glioblastoma: Together and/or Separately? International Journal of Molecular Sciences, 23(2):604 (2022).