KC-2697
MC38-BCMA-High-Cell-Line
Background of MC38-BCMA-High-Cell-Line
B cell maturation protein (BCMA), also named CD269, is a membrane of the TNF receptor super family(TNFRSF17) and mainly expressed in mature B-lymphocytes, and play an important role in B cell development and autoimmune response.
Specifications
| Catalog Number | KC-2697 |
|---|---|
| Cell Line Name | MC38-BCMA-High-Cell-Line |
| Host Cell Line | MC38 |
| Description | Stable MC38 cell line expressing exogenous BCMA gene in high level |
| 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 + 5μg/ml Puromycin |
| Selection Marker | Puromycin |
| Morphology | Epithelial |
| Subculture | Split saturated culture 1:4~1:8 every 2~3 days; seed out at about 1-2 × 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 | Yes |
Cell Line Generation
MC38 BCMA Cell Line was generated using a lentiviral vector expressing the BCMA sequence.
Characterization
Figure 1: Characterization of BCMA overexpression in the MC38 BCMA stable clone using FACS.
Figure 2: Validation of in vivo tumorigenicity of MC38 cells stably expressing high levels of BCMA via subcutaneous implantation in C57BL/6J mice, with tumor growth monitored by measuring volume (V=0.5×length×width²) and body weight.
Cell Resuscitation
1. Prewarm culture medium (DMEM supplemented with 10% FBS and 5μg/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 1-2 × 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 1-2 × 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. Gross, J. A. et al. TACI and BCMA are receptors for a TNF homologue implicated in B-cell autoimmune disease. Nature 404, 995–999 (2000).
2.Hatzoglou, A. et al. TNF Receptor Family Member BCMA (B Cell Maturation) Associates with TNF ReceptorAssociated Factor (TRAF) 1, TRAF2, and TRAF3 and Activates NF- B, Elk-1, c-Jun N-Terminal Kinase, and p38 Mitogen-Activated Protein Kinase. The Journal of Immunology 165, 1322–1330 (2000).
2.Hatzoglou, A. et al. TNF Receptor Family Member BCMA (B Cell Maturation) Associates with TNF ReceptorAssociated Factor (TRAF) 1, TRAF2, and TRAF3 and Activates NF- B, Elk-1, c-Jun N-Terminal Kinase, and p38 Mitogen-Activated Protein Kinase. The Journal of Immunology 165, 1322–1330 (2000).