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Cells were maintained in a humidified environment of 5% CO 2 at 37 C, cultured until an optical confluence of 75-100%, detached with 0.05% trypsin-EDTA 1 (25300-054, Invitrogen, Thermo Fisher Scientific, Waltham, Massachusetts, USA), at 37 C for approximately 5 min, and split to new plates according to the experimental procedures
chronic constipation, abdominal pain and constant extreme fatigue) and were refractory to multiple medications
Human clinical trials establishing these effects have not been completed and conclusions about efficacy in humans should not be drawn from this data alone
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