What is the Difference Between Warm and Cold Trypsinization?

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The main difference between warm and cold trypsinization lies in the temperature at which the trypsin enzyme is added for cellular disaggregation. Here are the key differences between the two methods:

  • Warm Trypsinization:
  • Carried out at 37°C (36.5 - 37°C).
  • Yields a lesser amount of viable cells due to the adverse effect of warm trypsin.
  • Involves incubation of tissue with warm trypsin.
  • May require centrifugation steps.
  • Cold Trypsinization:
  • Carried out under ice-cold conditions, typically at 4°C.
  • Yields a higher number of viable cells as cell damage is minimized.
  • Involves soaking the tissue with cold trypsin at 4°C prior to incubation with warm trypsin.
  • Minimizes cell damage and does not use centrifugation steps.

Both warm and cold trypsinization methods are used in the enzymatic disaggregation of cells in animal tissues for primary cell culturing. While warm trypsinization is more common, cold trypsinization has been found to have some advantages, such as a higher yield of viable cells and being less laborious. However, cold trypsinization may take longer to complete.

Comparative Table: Warm vs Cold Trypsinization

Warm and cold trypsinization are two methods used in enzymatic disaggregation of cells in animal tissue cultures. The main difference between these methods is the temperature at which the trypsin enzyme is added for cellular disaggregation. Here is a table summarizing the differences between warm and cold trypsinization:

Method Temperature Cell Viability Cell Damage Trypsin Solution Centrifugation
Warm Trypsin 36.5-37°C Lower High Warm Trypsin Intensive
Cold Trypsin 4°C Higher Low Cold Trypsin Less
  • Temperature: Warm trypsinization takes place at 36.5-37°C, while cold trypsinization occurs at 4°C.

  • Cell Viability: Cold trypsinization yields a higher number of viable cells due to minimized cell damage, while warm trypsinization results in a lower number of viable cells.

  • Cell Damage: Warm trypsinization causes more cell damage compared to cold trypsinization, which is less damaging to cells.

  • Trypsin Solution: Warm trypsinization uses warm trypsin, while cold trypsinization uses cold trypsin.

  • Centrifugation: Warm trypsinization involves intensive centrifugation, while cold trypsinization requires less centrifugation.