- To prepare individual tissue cells from a primary culture, the cell-cell and cell-matrix interaction must be broken. To achieve this would NOT use:
(1) EDTA (2) Trypsin
(3) Collagenase (4) SeparaseThe correct answer is (4) Separase.
Option-wise explanation
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(1) EDTA – used
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EDTA is a calcium/magnesium chelator.
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It disrupts cadherin- and integrin-mediated adhesion by removing divalent cations, helping detach cells from each other and from the matrix.
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Commonly used with or without enzymes when dissociating tissues or adherent monolayers.
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(2) Trypsin – used
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Trypsin is a serine protease that cleaves peptide bonds in extracellular matrix proteins and cell-surface adhesion molecules.
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Widely used to dissociate cells from tissue slices or culture flasks into single-cell suspensions.
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(3) Collagenase – used
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Collagenase specifically digests collagen, a major component of extracellular matrix.
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Especially important for primary tissues rich in connective matrix (liver, fat, cartilage), allowing embedded cells to be released.
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(4) Separase – NOT used (correct option)
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Separase is an intracellular cell-cycle protease that cleaves cohesin complexes during anaphase to separate sister chromatids.
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It acts inside the nucleus, not on extracellular matrix or adhesion proteins, and is not applied as a reagent for tissue dissociation.
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Therefore, the chemical/enzymatic tools appropriate for breaking cell–cell and cell–matrix interactions are EDTA, trypsin, and collagenase, while separase is unrelated to tissue dissociation, making option (4) the one that would NOT be used.
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