Trypsin-EDTA solution, 0,25%, bioReagent, suitable for cell culture.

• Sterile-filtered
• Combined of 2,5 g porcine trypsin and 0,2 g EDTA, 4Na per liter of Hanks′ balanced salt solution with phenol red
• Stored frozen between –10 and –40 °C, repeated cycles of freezing and thawing should be avoided

Trypsin consists of a single chain polypeptide of 223 amino acid residues, produced by the removal of the N-terminal hexapeptide from trypsinogen which is cleaved at the Lys - lle peptide bond. The sequence of amino acids is cross-linked by 6 disulfide bridges. This is the native form of trypsin, beta-trypsin. BETA-trypsin can be autolyzed, cleaving at the Lys - Ser residue, to produce alpha-trypsin. Trypsin is a member of the serine protease family.

Trypsin cleaves peptides on the C-terminal side of lysine and arginine residues. The rate of hydrolysis of this reaction is slowed if an acidic residue is on either side of the cleavage site and hydrolysis is stopped if a proline residue is on the carboxyl side of the cleavage site. The optimal pH for trypsin activity is 7 to 9. Trypsin can also act to cleave ester and amide linkages of synthetic derivatives of amino acids. EDTA is added to trypsin solutions as a chelating agent that neutralizes calcium and magnesium ions that obscure the peptide bonds on which trypsin acts. Removing these ions increases the enzymatic activity.

Trypsin-EDTA solution was used in detaching HT29 human colorectal cancer cells cultured in RPMI 1640 which was supplemented with 10% fetal calf serum, during relative cell frequency determination of high concentration samples. Trypsinize the transient transfected human embryonic kidney tcA-201 cell line. Enzymatically release mouse fibroblasts cells (cell line L929) adhered to the scaffold, during cell culturing to assess the influence of several modified treatments of Poly(L/D)lactide 96/4 non-woven scaffolds and fibres.