Disaggregation by Trypsin in Embryonic Tissue, A Trypsin Inhibitor, Trypsinization, Genes, Centrifugation, Disaggregation

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	Disaggregation by Trypsin in Embryonic Tissue Crude trypsin is the most common enzyme used for disaggregation for the following reasons: (i) it is tolerated by a variety of cells, (ii) it is effective for many tissues and (iii) its residual activity is neutralized by serum of the medium or by a trypsin inhibitor (e.g. soybean trypsin inhibitor) in case of a serum free medium.
To facilitate exposure of cells to the warm enzyme (36.5°C) for a minimum period, dissociated cells are collected every half an hour and the trypsin is removed by centrifugation after 3-4 hours required for complete disaggregation. Alternatively cold trypsinization may be used which involves soaking of tissue in trypsin at 40°C (to allow penetration of enzyme), followed by incubation at 36, 5°C for a shorter period. Different steps of warm trypsinization and cold trypsinization.