Biliary slices were transferred from the cold oxygenated KHB solution to prewarmed oxygenated 12‐well plates filled with 1.3 mL Williams’ medium E glutamax‐I (Gibco, Paisley, United Kingdom) containing 1.1 g/mL of D‐glucose and 2 mg/mL ciprofloxacin per well (Fig. 1 ). Slices at baseline were harvested directly after the slicing procedure. Plates with slices were placed in a shaking incubator (shaking at 90 cycles per minute at 37°C) under continuous supply of 80% O2 /5% CO2.9 Thereafter, every 24 hours, the medium was refreshed and slices were harvested for immunohistochemistry and for real‐time quantitative PCR for up to 144 hours. For immunohistochemistry, harvested slices were fixed in 10% buffered formalin and embedded in low‐temperature fusion paraffin (55°C‐57°C), and 3‐4‐µm sections were prepared and mounted on glass slides for histological and immunohistochemical assessment. For real‐time quantitative PCR, harvested slices were preserved in 400 mL of RNAlater solution (Sigma‐Aldrich, Steinheim, Germany), snap‐frozen, and stored at −80° C.
Williams medium e glutamax 1
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Williams' medium E glutamax‐I is a cell culture medium formulated to support the growth and maintenance of various cell types. It contains the amino acid glutamine, which is an essential nutrient for many cell lines. The medium is designed to provide a balanced and defined environment for cell cultivation.
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Lab products found in correlation
2 protocols using williams medium e glutamax 1
1
Culturing Precision-Cut Biliary Slices
2
Kidney Slice Incubation in Oxygenated Medium
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Kidney slices were washed quickly by gently transferring them into WME to remove the UW. Before adding medium and mPCKS, 12-well plates were coated with 10% bovine serum albumin (BSA) in milliQ water for 30 min. The solution was removed and plates were air-dried. Slices were then incubated in the coated plates containing 1.3 ml Williams Medium E glutamax-I (Gibco, Paisley, UK) per well, supplemented with 25 mM D-glucose and 50 µg/ml gentamycin. Medium was pre-warmed and gassed with 80% O2/5% CO2 before it was added to the wells. Slices were individually transferred to the wells of a culture plate placed on a surgical mattress to maintain the medium at 37°C. Plates were immediately transferred back in a shaking CO2 incubator (90 cycles/min) under continuous supply of 80% O2/5% CO2, as the pH of the medium rapidly changes in normal atmosphere. Slices were incubated for 1, 6, 12, 24, 48 or 72 h. When appropriate, after 24 and 48 h slices were transferred to new plates with fresh medium. For refreshing, new culture plates were filled with medium, and the plates were pre-warmed and oxygenated. The incubated plates and new plates were placed on a surgical mattress to maintain the temperature at 37°C. The slices were quickly transferred from the incubated plates into the new plates with a spatula and placed in the incubator.
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