Difficulty: Easy
Correct Answer: Releasing the cells from their supporting extracellular matrix without damaging viability
Explanation:
Introduction / Context:
Preparing primary cell suspensions from tissues (liver, kidney, tumors) requires breaking cell–cell junctions and cell–matrix adhesions while preserving cell viability and functionality. This is foundational for primary culture, single-cell analyses, and regenerative studies.
Given Data / Assumptions:
Concept / Approach:
The major challenge is to release cells from the ECM and from each other without excessive shear or proteolytic damage. Enzymes such as collagenase, dispase, elastase, and trypsin, plus chelators (EDTA/EGTA) and mechanical trituration, are used carefully to disrupt ECM and adhesions. Over-digestion or harsh shear compromises viability and surface receptors.
Step-by-Step Solution:
Verification / Alternative check:
Quality checks include trypan blue viability, yield counts, and functional assays; optimizing enzyme concentration, temperature, and time improves outcomes.
Why Other Options Are Wrong:
Common Pitfalls:
Overexposure to proteases, insufficient washing, and excessive shear leading to low viability or altered phenotype; ignoring tissue-specific protocols.
Final Answer:
Releasing the cells from their supporting extracellular matrix without damaging viability
Discussion & Comments