ITS Supplement (Insulin-Transferrin-Selenium)
Quick Facts
| Feature | Information |
|---|---|
| Category | Cell Culture Supplement |
| Type | Chemically defined serum replacement |
| Typical Stock | 100× |
| Typical Working Concentration | 1× |
| Primary Function | Supports serum-free and reduced-serum cell culture |
| Common Basal Media | DMEM/F-12, Williams' E, DMEM, IMDM |
| Common Applications | Stem cells, hepatocytes, epithelial cells, organoids, primary cells |
Overview
ITS Supplement is a chemically defined mixture of Insulin, Transferrin, and Selenium that replaces several growth-supporting functions normally provided by fetal bovine serum (FBS). It promotes cell survival, metabolism, and proliferation while reducing variability associated with serum-containing media.
ITS is widely used in serum-free and reduced-serum culture systems, particularly for stem cells, hepatocytes, epithelial cells, endothelial cells, and organoid cultures.
Composition
| Component | Function |
|---|---|
| Insulin | Stimulates glucose uptake, metabolism, and protein synthesis |
| Transferrin | Delivers iron required for cell growth and DNA synthesis |
| Selenium | Supports antioxidant enzymes and protects against oxidative stress |
Note: Commercial formulations (e.g., ITS-A, ITS-G, ITS-X) may contain additional components or modified concentrations. Always consult the manufacturer's specifications.
Biological Role
ITS replaces several essential serum functions while maintaining a defined culture environment.
- Insulin promotes glucose uptake, amino acid transport, and activation of anabolic signaling pathways.
- Transferrin supplies bioavailable iron while minimizing oxidative damage from free iron.
- Selenium functions as a cofactor for antioxidant enzymes, supporting cellular redox balance and viability.
Together, these components improve growth and reproducibility while minimizing the variability introduced by serum.
Common Applications
ITS is commonly used for:
- Serum-free and reduced-serum culture
- Primary hepatocyte maintenance
- Stem cell differentiation
- Mesenchymal stem cell culture
- Endothelial and epithelial cell culture
- Liver and brain organoids
- Tissue engineering
- Developmental biology
- Drug screening
Common Cell Types
| Cell Type | Typical Application |
|---|---|
| Primary Hepatocytes | Serum-free maintenance |
| Human ESCs | Directed differentiation |
| Human iPSCs | Differentiation protocols |
| Mesenchymal Stem Cells | Expansion |
| Endothelial Cells | Reduced-serum culture |
| Epithelial Cells | Defined media |
| Fibroblasts | Reduced-serum culture |
| Brain Organoids | Neural differentiation |
| Liver Organoids | Hepatic maturation |
Typical Media Formulations
Primary Hepatocytes
- Williams' E Medium
- ITS Supplement
- Dexamethasone
- GlutaMAX
Stem Cell Differentiation
- DMEM/F-12
- ITS Supplement
- Growth factors (cell type dependent)
Liver Organoids
- Williams' E Medium
- ITS Supplement
- HGF
- Oncostatin M
Endothelial Cells
- DMEM
- ITS Supplement
- Endothelial growth supplements
Advantages
- Chemically defined composition
- Reduces dependence on serum
- Improves experimental reproducibility
- Supports cellular metabolism
- Enhances antioxidant protection
- Compatible with many primary cell types
- Suitable for serum-free media development
Limitations
- Does not completely replace serum for all cell types
- Additional growth factors are often required
- Performance depends on the specific cell type and protocol
- Commercial formulations are not always interchangeable
Comparison with Related Supplements
| Feature | ITS | FBS | KSR |
|---|---|---|---|
| Composition | Defined | Undefined | Semi-defined |
| Batch Variability | Low | High | Low |
| Stem Cell Maintenance | Moderate | Moderate | Excellent |
| Hepatocyte Culture | Excellent | Good | Limited |
| Serum Replacement | Partial | Complete serum | Stem cell specific |
References
- Bottenstein JE, Sato GH. Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proceedings of the National Academy of Sciences (1979).
- Freshney RI. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications.
- Gibco™ ITS Supplement Product Information.
- Thermo Fisher Scientific Cell Culture Handbook.
- ATCC Animal Cell Culture Guide.