0.25% Trypsin-EDTA Solution 1X
In adherent cell culture, trypsin or trypsin-EDTA (ethlyenediaminetetraacetic acid) is used for the detachment of cells from a culture dish or flask, and degradation of cell-cell adhesion. Trypsin, a proteolytic enzyme, can cleave collagen that is pre-coated on culture surfaces for cell adhesion. EDTA, a chelating agent, can bind with cations such as Ca2+ and Mg2+ which are involved in cell adhesion. Subculture usually requires chelation of cations and proteolytic reactions, but the severity of the treatment (high concentration or time length) will reduce the viability and the rate of cell growth because of the over-degradation of cell surface proteins and deficiency of cations essential for cells. A protocol should be selected with the least severity possible while ensuring it is compatible with the generation of a single cell suspension of high viability.
PTS-BTG-XXX-000 contains 0.25% (w/v) gamma irradiated porcine trypsin (PTR-BTG-XXX) and 0.9 mM EDTA·4Na.2H2O in Hanks’ balanced salt solution (HBSS). PTS-BTG-XXX-000 is sterile filtered, tested for porcine parvovirus, and is mycoplasma and cell culture tested.
The concentration of Trypsin-EDTA is strongly influenced by cell type, culture type, and characteristics of cell growth. It is important to review the literature for recommendations concerning medium, supplementation and physiological parameters required for a specific cell line.
Trypsin-EDTA solutions should be kept at ≤ -10°C in the dark for long term storage, but can be stored at 2-8°C for one week. Deterioration of the solution may be recognized by:
(1) Precipitate or particulate matter throughout the solution
(2) Cloudy appearance
(3) Color change
(4) pH change
The nature of supplements added may affect storage conditions and shelf life of the medium. Product label bears expiration date.
Biological Performance Characteristics
The detachment and growth-maintaining capacities of Trypsin-EDTA are tested in a medium supplemented with 10% FBS using appropriate adherent cell lines.
For In Vitro Use Only
|Appearance||Visual||Red Transparent Solution|
|pH at RT Before Additions||pH Meter||7.0 - 8.0|
|Osmolality||Chemistry Analyzer||265 - 315 mOsm/kg H2O|
|Gamma Irradiation||25 - 40 kGy|
|Sterility||Sterilized by 0.2 um filtration system|
|Porcine Trypsin||0.25% (w/v)|