Effect of Trypsinisation Time on Cell Viability and Dissociation Efficiency in Chick Embryo - Derived Cells

AbstractEnzymatic dissociation is a biological process that uses enzymes to break down the extracellular matrix and separate cells from tissues. It is an important step in the formation of primary cell cultures. However, optimizing the exposure time is essential to ensure efficient cell separation. In this study, 96-hour old chick embryo tissue was subjected to 0.25% trypsin and 1mM EDTA treatment for varying durations (2, 4, 6 and 8 minutes) to assess the effect of exposure time on cell dissociation and viability. Morphological analysis and Trypan Blue exclusion assay were performed to evaluate dissociation and survival of the cells. The results indicated that trypsinisation for two minutes led to incomplete tissue dissociation with minimal cell damage, while exposure for four minutes resulted in optimal cell separation and maximum viability., In contrast, treatment for six and eight minute durations showed a progressive decline in cell viability, with significant cell death observed at eight minutes. These findings indicate the importance of time-optimized trypsinisation in primary culture protocols. and Chick embryo-derived tissue exposure for 96-hour with 0.25% trypsin and 1mM EDTA for four minutes was found to be most effective for achieving balance between cell yield and viability.