An efficient and simple method of non-enzymatic detachment of monolayer cultures into single cell suspension
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Activating the Na+/H+ antiporter causes a distinctive and rapid endocytosis which internalizes the plasma membrane leading to cell retraction and detachment as rounded cells with much reduced surface area. Antiport activation is by a combination of two individually effective motivations, viz. a) steep [Na+] and [H+] gradients across the plasma membrane and b) allosteric activation by second messengers initiated with simple inorganic sulphate. The dissociated cells are as viable as those released by trypsinization. Thus it provides an effective enzyme-free alternative to trypsin digestion in cell dissociation.
Key wordscell dissociation detachment rounding Na+/H+ antiport endocytosis sulphate
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