Isolation of single-cell suspensions holds great value in biological and medical research, especially regarding cell analysis, drug testing, and tissue engineering. This involves detaching single cells from a culture surface and separating them from each other. EDTA and trypsin are the two critical reagents required to perform this task. Here, a blog will analyze these two reagents and how they facilitate cell dissociation, synergism in isolating a single cell.
What are EDTA and Trypsin?
EDTA is a chelating agent which bonds with divalent ions like calcium and magnesium which are very important for cell adhesion as they stabilize proteins on the surface of cell membranes to the extracellular matrix. When chelating these ions, EDTA weakens the bonds between the cells and the surfaces, thus permitting easy removal of the cells from the culture surfaces.
Trypsin is a protease enzyme that digests the proteins within a cell assembly. It cleaves extracellular matrix components such as fibronectin and laminin; this action allows the disconnection of cells from each other and from the surface they are attached to.
The role of EDTA in cell culture
EDTA typically acts in conjunction with other forms of dissociation in cell culture. It is used mainly for dismantling-capture of those vital ions of the organism that hold cells aggregated. As EDTA differs from both culture surface and cells, this often gives a very suitable environment for trypsin to carry out its job and therefore makes it very much the first in terms of critical ion for isolating cells from adherent culture surfaces.
Trypsinization for Isolating a Single-Cell Suspension
Trypsin is the primary enzyme in cell detachment once EDTA has weakened cell-cell and substrate-cell adhesion. It further degrades proteins, which manage the aforementioned mechanisms. Trypsin cleaves those proteins such that single cells can be freed from a surface and suspended in media. One has to watch over trypsin activity because sometimes, over-digestion leads to damaging effects regarding cell viability.
EDTA plus Trypsin
The combination of EDTA and trypsin is the most remarkable technique for obtaining one-pot viable single-cell suspensions. EDTA softens the adhesion, and thus, trypsin can more efficiently rupture the proteins holding the cells together. This mechanism makes EDTA trypsin treatment the gold standard for dissociating adherent cell cultures.
End Note
Both EDTA in cell culture and trypsin have very important roles in the isolation of single-cell suspensions for research. Knowing how to apply both very well forthwith gives researchers an assurance that they would be able to isolate functional, viable cells within the experiments.
Connect with PurMa Biologics LLC, if you are seeking quality reagents for your research.
