Cat.No. : CSC-SC015139-1 Host Cell: HEK293
| Cat. No. | CSC-SC015139-1 |
| Description | This cell line is engineered to stably express human steroid 5 alpha-reductase 2 (SRD5A2) in HEK293 cells. |
| Gene | SRD5A2 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| 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 |
The enzyme steroid 5-α reductase (5AR) converts steroids into 5-α reduced metabolites, such as 5-α dihydrotestosterone (DHT) from testosterone. Researchers developed a cell-based assay to evaluate the inhibition of 5AR, crucial for DHT production. IC50 values for inhibitors finasteride and dutasteride were determined using human and fish cell lines via transient overexpression. Fish cell lines showed a lower IC50 for dutasteride than human cell lines, suggesting species-specific responses. This sensitive assay, employing chemical derivatization and LC-MS/MS, exhibits high reproducibility and is suitable for screening endocrine-disrupting chemicals (EDCs). Future studies will explore 5AR inhibition by EDCs across species to elucidate their impact on endocrine pathways.
Figure 1. DHT levels were measured in non-vector-, SRD5A1-HEK293, and SRD5A2-HEK293 cells following testosterone addition. KM values reflecting enzyme affinity were calculated, followed by inhibition assays with finasteride and dutasteride to determine IC50 values (Kim D, et al., 2021).
A: The role of the Human SRD5A2 Stable Cell Line - HEK293 in studying androgen metabolism is to provide an experimental platform to investigate how the SRD5A2 enzyme catalyzes the conversion of testosterone to dihydrotestosterone, a process crucial for the male reproductive system and prostate health. This cell line helps elucidate the role of the SRD5A2 gene in androgen-related diseases, such as prostate cancer and benign prostatic hyperplasia.
A: The role of the Human SRD5A2 Stable Cell Line - HEK293 in studying androgen receptor signaling is to investigate how the stable expression of the SRD5A2 enzyme affects the activation and signaling of androgen receptors. This cell line helps reveal the role of SRD5A2 in regulating androgen signals and cellular functions, which is crucial for understanding the molecular mechanisms of androgen-related diseases.
A: The role of the Human SRD5A2 Stable Cell Line - HEK293 in identifying potential therapeutic targets for androgen-related diseases is to provide an experimental model for evaluating the impact of SRD5A2 enzyme inhibitors on disease progression. This cell line aids in discovering new therapeutic strategies, especially in the development of treatments for prostate cancer and benign prostatic hyperplasia.
A: The role of the Human SRD5A2 Stable Cell Line - HEK293 in studying the effects of androgens on cell proliferation is to simulate the role of androgens in cell growth and differentiation by stably expressing the SRD5A2 enzyme. This cell line helps understand the function of SRD5A2 in regulating cell growth and maintaining tissue homeostasis, which is significant for developing treatments that inhibit excessive cell proliferation.
A: The stable expression of the SRD5A2 gene in the Human SRD5A2 Stable Cell Line - HEK293 is achieved by integrating the SRD5A2 gene into the genome of HEK293 cells, using selective markers and screening strategies. This typically involves the use of viral vectors, such as retroviruses or lentiviruses, and antibiotic resistance genes as selection markers, thereby creating a cellular model that continuously expresses the SRD5A2 protein.
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Provides a platform for developing therapeutic drugs targeting SRD5A2-related diseases.
Contributes to understanding the association between SRD5A2 gene mutations and disease development.
The Human SRD5A2 Stable Cell Line - HEK293 aids in studying the role of the SRD5A2 gene in hormone metabolism.
Enhances knowledge of related physiological processes by studying the function of SRD5A2 within cells.
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