non-mammalian and non-bacterial based
Accutase™ is a ready to use non-mammalian, non-bacterial replacement for all applications of trypsin. It is a natural enzyme mixture with proteolytic and collangenolitic activities. It mimics the action of trypsin and collagenase simultaneously. However, because the enzymes operate more efficiently than trypsin and collagenase, it results in gentler and more effective cell detachment.
A few cell lines that Accutase™ has been shown to detach without harm:
|keratinocytes||vascular endothelial cells|
|vascular smooth muscle cells||hepatocytes|
|hepatocyte progenitors||primary chick embryo neuronal cells|
|bone marrow stem cells||adherent CHO cells|
|adherent BHK cells||macrophages|
|293 cells||L929 cells|
|immortalized mouse||testicular germ cells|
|M24 and A375 metastatic melanoma||gliomas U251 and D54|
|HT1080 fibrosarcoma cells||Sf9 insect cells|
Human MG63 Fibrosarcoma cells cultured in DMEM + 10% FBS were treated with Accutase™. Treatment resulted in rapid cell detachment, a single cell suspension, and high viability. Accutase is gentle on cells; viability was 97 ± 3% even after 45 minutes in Accutase.
Accutase performs exceptionally well in detaching cells for:
|hESC culturing||virus growth assay|
|analysis of cell surface markers||cell proliferation|
|tumor cell migration assays||routine cell passage|
|flow cytometry||quiescence assays by serum starvation|
|transformation assays by oncogene||transfectionneural crest cell migration assays|
|production scale-up (bioreactor)|
Accutase is formulated at a concentration that is ready to use, once defrosted. (Note: Never defrost a bottle of Accutase at 37°C.) A defrosted bottle of Accutase can be removed from the refrigerator and immediately applied to cells. It does not need to be and should not be pre-warmed to 37°C. Accutase contains proteolytic and collagenolytic enzymes to gently break down the cell adhesion structure on the outside of cells that attaches them to the bottom of the flask.
This entire procedure should be done in a laminar flow hood using proper aseptic technique.