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Biotase
Biotase - new trypsine alternative
To sub-culture cells, Trypsin derived from bovine, or porcine pancreas is still widely used in both academic, and industrial applications. There is endless supply of Trypsin from the slaughterhouses, and cell culturists do have a long experience in working with this product on different activity levels. However, some concern is raised through the fact that bovine derived material can possibly be BSE contaminated, and porcine material might have a certain load of parvovirus, indigenous to swine. In general, delicate applications do not allow for any bovine, or porcine, or any other vertrebrate derived material. Since a couple of years, there is an alternative available in the market, marketed as Accutase®, via a company called Innovative Cell Technologies.
In order to offer an alternative to porcine, or bovine derived disintegration enzyme, we have approached Innovative Cell Technologies for the patent protected raw material for further manufacturing. According to the information we have, this enzyme is processed from a marine invertebrate (we suspect it is a by-product of the crab meat industry). The non-vertebrate origin makes it a good alternative though, and its way of enzymatic cell separation is gentlier, and easier to control. In general, Biotase acts slowlier than any other disintegration enzyme with more cells surviving. The protocol is very simple:
- remove culture media
- wash cells using PBS without Ca/Mg (cat.no. L 1825)
- following removal of the buffer, add Biotase (5 ml per 75 cm2), warmed up at 37°C
- allow for 5 – 15 min (at 37°C) for digestion
- perform cell counting, and sealing of cell into new medium
Enzymatic activity of Biotase is blocked through the addition of Fetal Bovine Serum (FBS); however, in most cases this might not be necessary.
Repeated freezing and thawing cycles have to be avoided, smaller aliquots should be re-frozen. Once thawed, Biotase keeps its enzymatic activity up to 7 days, if stored at +2 to +8°C.
Biotase comes in a ready-to-use form (100 ml bottle, cat.no. L 2193). As with all newly introduced products, the labelled shelf life of Biotase is 12 months; we will expand this to possibly 36 months, if data allow to do so.
To sub-culture cells, Trypsin derived from bovine, or porcine pancreas is still widely used in both academic, and industrial applications. There is endless supply of Trypsin from the slaughterhouses, and cell culturists do have a long experience in working with this product on different activity levels. However, some concern is raised through the fact that bovine derived material can possibly be BSE contaminated, and porcine material might have a certain load of parvovirus, indigenous to swine. In general, delicate applications do not allow for any bovine, or porcine, or any other vertrebrate derived material. Since a couple of years, there is an alternative available in the market, marketed as Accutase®, via a company called Innovative Cell Technologies.
In order to offer an alternative to porcine, or bovine derived disintegration enzyme, we have approached Innovative Cell Technologies for the patent protected raw material for further manufacturing. According to the information we have, this enzyme is processed from a marine invertebrate (we suspect it is a by-product of the crab meat industry). The non-vertebrate origin makes it a good alternative though, and its way of enzymatic cell separation is gentlier, and easier to control. In general, Biotase acts slowlier than any other disintegration enzyme with more cells surviving. The protocol is very simple:
- remove culture media
- wash cells using PBS without Ca/Mg (cat.no. L 1825)
- following removal of the buffer, add Biotase (5 ml per 75 cm2), warmed up at 37°C
- allow for 5 – 15 min (at 37°C) for digestion
- perform cell counting, and sealing of cell into new medium
Enzymatic activity of Biotase is blocked through the addition of Fetal Bovine Serum (FBS); however, in most cases this might not be necessary.
Repeated freezing and thawing cycles have to be avoided, smaller aliquots should be re-frozen. Once thawed, Biotase keeps its enzymatic activity up to 7 days, if stored at +2 to +8°C.
Biotase comes in a ready-to-use form (100 ml bottle, cat.no. L 2193). As with all newly introduced products, the labelled shelf life of Biotase is 12 months; we will expand this to possibly 36 months, if data allow to do so.