Cat.No. : CSC-SC004980-1 Host Cell: HEK293T
| Cat. No. | CSC-SC004980-1 |
| Description | This cell line is engineered to stably express human epithelial cell adhesion molecule (EPCAM) in HEK293 cells. |
| Gene | EPCAM |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HEK293T |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Studying the interactions between immune cells and cancer cells 2. Studying the mechanisms of resistance to immune checkpoint blockade 3. High-throughput screening 4. Drug target validation |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:3-1:6. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
Epithelial cell adhesion molecule (EpCAM) has been shown to be strongly expressed in human breast cancer and cancer stem cells, and its overexpression is thought to support tumor progression and metastasis. Here, researchers analyzed the effects of transient overexpression of EpCAM by adenovirus on proliferation, migration, and differentiation of primary human mammary epithelial cells (HMECs). Overexpression of EpCAM in HMECs did not significantly alter the gene expression profile of proliferating or growth-arrested cells. Proliferating HMECs displayed predominantly glycosylated EpCAM isoforms and inhibited cell proliferation and migration through upregulation of p27KIP1 and p53. EpCAM-overexpressing HMECs showed downregulation of E-cadherin. Furthermore, cells were more resistant to TGF-β1-induced growth arrest and maintained their ability to proliferate in vitro for a longer period of time. Xenografts of HMECs overexpressing EpCAM in chick embryos exhibited hyperplastic growth, deficient tube formation, and increased infiltration of chick leukocytes. Thus, EpCAM is a prosurvival factor that counteracts terminal differentiation processes in the normal mammary gland.
Compared to HMEC, EpCAM-overexpressing MCF10A showed increased cell proliferation (Figure 1B) and upregulated c-myc gene expression (Figure 1C). Changes in c-myc expression could also be monitored at the protein level (Figure 1D/E). In addition, MCF10A cell lines were generated by a lentiviral system with stable expression of either a non-silencing control (ns/crtl) or EpCAM-specific (E#2) shRNA. Both cell lines, MCF10A ns/crtl and MCF10AE#2, were transfected by adenovirus to overexpress GFP or EpCAM/GFP. MCF10AE#2 cells significantly downregulated EpCAM transcript levels at 24 and 48 h after adenovirus transfection compared to MCF10A ns/crtl cells (Figure 1F). After adenoviral EpCAM overexpression, real-time cell proliferation of MCF10AE#2 cells was significantly lower than that of MCF10Ans/crtl cells (Figure 1G). These data clearly demonstrate that EpCAM overexpression can enhance proliferation and c-myc levels in immortalized human mammary epithelial cells.
Figure 1. EpCAM overexpression leads to c-myc upregulation and increases cell proliferation in immortalized MCF10A human breast epithelial cells. (A) Flow cytometric analysis of EpCAM-overexpressing MCF10A cells. (B) Overexpression of EpCAM leads to a significant increase in cell proliferation under serum-reduced conditions. (C) Overexpression of EpCAM upregulates c-myc gene expression. (D) Western blot analysis of EpCAM overexpression and upregulation of c-myc protein levels. (E) Densitometry analysis of the ratio of c-myc to tubulin. (F) MCF10AE#2 cells significantly downregulate EpCAM transcript levels at 24 and 48 h after adenoviral transfection compared to MCF10Ans/crtl cells. (G) Real-time cell proliferation of MCF10AE#2 cells after adenoviral EpCAM overexpression is significantly lower than that of MCF10Ans/crtl. (Martowicz A, et al., 2013)
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The expression level of EPCAM in this stable cell line is very high. This significantly improves the sensitivity and accuracy of our assays, allowing us to obtain clear and conclusive experimental data.
The Human EPCAM Stable Cell Line - HEK293 is very easy to use. The cells are easy to culture and handle, which simplifies our workflow and makes experimental setup faster and more efficient.
The viability of these HEK293 cells is excellent. Even after multiple passages, the cells remain healthy and strong, which is essential for long-term studies and ensures the integrity of my research.
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