Initiating a primary cell culture from a surgical tissue is often difficult because of contamination. It is therefore important to incubate the tissue in an appropriate solution containing an optimized mixture of antibiotics, each at a given concentration able to avoid infectious contamination without affecting cell viability. Furthermore, it is not always possible to initiate primary cell culture immediately after surgical excision so the tissue needs to be stored for several hours until further processed.
Product Description
InfectoSTOP (GENT 19 - INSTOP) is a ready-to-use solution containing an optimized mix of antibiotics against gram-negative and gram-positive bacteria, mycoplasma and fungi for cell culture applications.
Simply dilute the necessary volume (enough to cover the tissue) of InfectoSTOP in PBS or in cell culture medium.
Intended Use
InfectoSTOP is intended for use in primary cell culture initiation or to extend cell viability in fresh surgical material of any tissue type.
InfectoSTOP can also be used for decontaminating established cell cultures.
The product should be handled under sterile conditions and is not intended for animal or human use.
Caution: If handled improperly, some components of this product may present a health hazard. Take appropriate precautions when handling this product, including the wearing of protective clothing and eyewear. Dispose of properly
How to use InfectoSTOP
To initiate primary cultures from fresh surgical material
Dilute InfectoSTOP 10x in PBS (4 °C) and put the tissue in the freshly made solution. Rinse tissue immediately twice or more and then incubate 2 hours at 4 °C. Prepare a fresh solution, rinse once and incubate overnight (or at least one week without viability loss, depending on the type of tissue) at 4 °C.
Just before initiating primary cell culture, rinse one more with InfectoSTOP.
To eliminate contamination in anchorage-dependant cells
- - Entirely rinse the flask twice with a 5x diluted fresh made InfectoSTOP solution (w/o serum)
- - Incubate at 37 °C for 1 hour
- - Entirely rinse the flask twice with a 10x diluted freshly made InfectoSTOP solution (w/o serum)
- - Incubate at 37 °C for 3 hours
- - Entirely rinse the flask twice with a 10x diluted freshly made InfectoSTOP solution (w/o serum)
- - Incubate overnight in cell culture with a 10x diluted freshly made dilution of InfectoSTOP in complete culture medium
The above cell treatment can be repeated twice if necessary.
The dilution and incubation time of InfectoSTOP must be adjust to your own cell culture
To eliminate contamination in anchorage-independent cells
Same procedure as above, but before rinsing and incubation, pellet the cells by brief centrifugation.
Storage and Stability
- InfectoSTOP is stored under -20° C at our facility and is shipped on dry ice.
- If the product is to be used immediately, thaw in a 37° C water bath or overnight at 4° C.
- If thawed in a water bath, do not leave the product at 37° C for more than 1 hour.
- When stored at 4° C, InfectoSTOP is stable for at least 2 weeks.
- If the product is not to be used within 1 week after receipt, we recommend storing it at below -20° C in a freezer that is not self-defrosting.
- Do not thaw and refreeze more than once. When stored at below -20°C, the product is stable until the expiration date shown on the label.
Handling:
GLP techniques should be employed for the safe handling of this product.
This includes observing the following practices:
- - Wear appropriate laboratory cloches including a lab coat, gloves and safety glasses.
- - Do not mouth pipette, inhale, ingest or allow to come into contact with open wounds. Wash thoroughly any area of the body, which comes into contact with the product.
- - Avoid accidental autoinoculation by exercising extreme care when handling in conjunction with any injection device.
- - Handle the product under sterile area.
- - This product is intended for research purposes and should be handled by qualified personnel only. It is not intended for use in humans or in animals. Gentaur is not liable for any damages resulting from the misuse or handling of this product.
Components
One bottle of 25 ml.