In the world of cell culture, detaching adherent cells from surfaces and each other without harming them is essential for countless in vitro applications. This is where our specially formulated solution of 0.25% Trypsin and 1mM EDTA in Hank’s Balanced Salt Solution comes in, acting as a gentle but effective tool for disaggregating cells with precision.
What’s Inside?
- 0.25% Trypsin (1:250): Trypsin is a proteolytic enzyme which cleaves the protein peptide bonds that help adherent cells stick to surfaces. At a 0.25% concentration, this formulation is just strong enough to efficiently detach cells from the substrate while remaining gentle enough to avoid damaging their structure. This solution uses trypsin 1:250, which, at full strength, can digest 250 parts casein for every one part trypsin.
- 1mM EDTA: Ethylenediaminetetraacetic acid (EDTA) is a chelating agent which binds the calcium and magnesium ions that help cells stick to each other. The 1mM concentration in this formulation sequesters just enough of these ions to improve cell disaggregation without disrupting the cell membrane.
- Hank’s Balanced Salt Solution: HBSS provides a stable, balanced environment for cells during the detachment process by maintaining the pH and osmotic balance of the solution. It also provides the cells with water and inorganic ions, ensuring they stay healthy as they are released from their adherent bonds.
- No Calcium or Magnesium: Calcium and magnesium are purposely excluded, as they help cells stick to surfaces and each other. Without these ions, EDTA and trypsin work more effectively.
Tested for Safety
This solution is rigorously tested to ensure it is free of both porcine parvovirus and mycoplasma contamination, which could otherwise compromise cell health. Combined with low levels of endotoxin and assured efficacy, researchers can confidently use this solution without risking contamination of their cell lines.
How It’s Used
This Trypsin-EDTA Solution is typically applied to cell culture plates or flasks to detach adherent cells. Here’s a quick look at the process:
- Remove Depleted Media: The adherent cell growth media is carefully removed and discarded.
- Rinse the Cells: Cells are usually rinsed with a balanced salt solution to remove any residual media that could interfere with trypsin.
- Disassociate the Cells: A small amount of Trypsin-EDTA in HBSS is added to the cells and incubated briefly, allowing the trypsin and EDTA to gently break down the proteins and ions that anchor the cells. Within a few minutes, the cells will have detached from the surface and disaggregated from each other
- Neutralize the Trypsin: Once the cells are disassociated, the trypsin is neutralized to prevent the proteolytic reaction from continuing and damaging the cells. This is done by adding fetal bovine serum, contained in either a trypsin neutralization solution or in the new adherent cell growth medium.
Applications in Research
Trypsin solutions like this one are invaluable for researchers working with adherent cell lines. They are used in:
- Passaging cells: Detaching cells to transfer them into fresh culture media for continued growth.
- Transfection experiments: Separating one or multiple cells in order to introduce foreign nucleic acids into their cytoplasm(s).
- Single-cell analysis: Isolating individual cells for detailed study.
In Summary
For laboratories handling adherent cell lines, having a safe, reliable and effective method to dissociate and resuspend adherent cells is crucial. Our 0.25% Trypsin and 1mM EDTA in Hank’s Balanced Salt Solution is expertly designed to optimize cell dissociation while preserving viability. With careful formulation and rigorous testing, researchers can trust this solution to deliver consistent, high-quality results across a wide range of applications, from routine passaging to complex experiments. Optimize your cell culture processes and elevate your research outcomes with our Trypsin-EDTA Solution today!
