Introduction
When carrying out cellular biology research, the culturing of adherent cells under optimal conditions is a central drive. Biotech researchers are increasingly using methods that promote cell health and functionality while minimizing reliance on animal-derived products, such as sera. Defined Bioscience's HiDef-ITS (Insulin, Transferrin, and Selenium) supplement can help meet these goals. Designed to improve the routine maintenance and growth of a broad spectrum of cell types, HiDef-ITS offers a robust formulation to support scientific advancements while minimizing or eliminating reliance on animal products.
The Development of HiDef-ITS
The use of insulin, transferrin and selenium for cell culture has been long recognized as helpful for serum-free growth of stem cells.1, 2 Addition of HiDef-ITS to our growing catalog was driven by a clear need within the scientific community for a more defined, reliable, and ethical approach to cell culture. In addition to ethical concerns regarding animal sourcing, traditional serum-based media, while effective, pose several scientific challenges, including batch-to-batch variability, potential interference from undefined components, and the possibility of contamination or prions. These issues often lead to inconsistencies in experimental results and complicate the scaling of cell-based applications.
Recognizing these challenges, Defined Bioscience sets out to formulate serum replacements that not only match but exceed the performance of traditional fetal bovine serum (FBS). HiDef-ITS is a defined, animal origin-free (AOF), and feeder-free supplement that reduces or eliminates the use of FBS for many cell types, thereby enhancing the reproducibility and reliability of cell culture experiments. Unlike other serum replacement solutions on the market, HiDef-ITS is made with entirely animal origin-free componentry. .
Composition and Benefits
HiDef-ITS combines three critical components, each selected for their specific roles in supporting cell culture:
Insulin: Insulin is crucial for promoting glucose uptake, amino acid transport, and overall cell metabolism, which are vital for cell growth and proliferation.
Transferrin: As a high-affinity iron carrier, transferrin not only facilitates iron transport but also acts as an antioxidant, reducing the oxidative stress on cells that can lead to premature senescence or apoptosis.
Selenium: As sodium selenite, this component acts both as an antioxidant and as a necessary cofactor for various seleno-enzymes involved in cellular defense mechanisms.
These components are provided in a concentrated 100x format, dissolved in Earle’s Balanced Salt Solution (EBSS) without phenol red, adhering to Defined Bioscience’s commitment to using chemically defined and animal free componentry. This formulation ensures that cells receive necessary nutrients and protective agents without the variability introduced by serum.
Practical Applications and Versatility
HiDef-ITS is designed for use across a wide range of cell types, including primary cells, immortalized cell lines, and stem cells. It is particularly effective in environments where low-density attachment and reduced serum conditions are required. The supplement can dramatically reduce or eliminate the requirement for FBS without compromising cell health or experimental outcomes. HiDef-ITS is recommended for use at 1x concentration (a 1:100 dilution of the provided 100x stock), but has been used at lower and higher dilutions depending on the user’s needs and applications.
This versatility makes HiDef-ITS ideal for use in a variety of scientific and medical research fields, including toxicology, drug discovery, and regenerative medicine. By maintaining cells in a more defined and controlled environment, HiDef-ITS facilitates more accurate and reproducible results in research and therapeutic development applications.
User Experience and Recommendations
At Defined Bioscience, we prioritize not only the performance of our products but also the ease of their integration into existing laboratory workflows. HiDef-ITS is simple to use:
Storage and Preparation: The supplement should be stored at 2-8°C and protected from light, with a shelf life of eighteen months from the date of manufacture. It is easily activated by warming to room temperature before use. We do not recommend warming HiDef-ITS above ambient before use, for example in a heat bath or incubator.
Application: HiDef-ITS should be added to a basal nutrient medium, such as our DMEM/F12 with stabilized ւ-alanyl-ւ-glutamine, as per the specific requirements of the cell type being cultured. Adjustments in FBS concentrations and other supplements should be made based on the particular needs of the experiment.
Conclusion
HiDef-ITS by Defined Bioscience provides a defined, animal-free, effective, and versatile serum replacement. Whether for basic research or complex biotechnological applications, HiDef-ITS enhances the consistency, reliability, and ethical standards of cell culture practices.
We encourage researchers looking for a defined approach to cell culture to consider integrating HiDef-ITS into their laboratory protocols. By doing so, they will not only improve the quality of their cell cultures, but also contribute to the broader scientific endeavor of creating more sustainable and reproducible research methodologies.
References
- Mather JP. Making informed choices: medium, serum, and serum-free medium. How to choose the appropriate medium and culture system for the model you wish to create. Methods Cell Biol. 57:19-30 (1998). doi: 10.1016/s0091-679x(08)61569-1
- Ludwig T, Bergendahl V, Levenstein M, et al. Feeder-independent culture of human embryonic stem cells. Nat Methods 3:637–646 (2006). doi: 10.1038/nmeth902
- HiDef-ITS Product Guide