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Begin machining into the chamber to a depth of 2.84 mm while moving along the y-axis (positive direction) to 7.62 mm (0.300") and then traverse to 7.62 mm (0.300") in the opposite (negative) direction to a complete reservoir length of 15.24 mm (0.600"). Offset the bit to 3.03 mm ...
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48 ml of supernatant was centrifuged at 75,500 × gfor 1.5 h. After centrifugation, 20–30 μl of DMEM was left in the tube to dissolve the pellet containing RIA. The resulting viral stock has a titer of 1 × 107 pfu/ml. Viral stocks were stored in −80 °C...
(Cat#356231, Corning)-coated plates, cultured in mTeSR1 complete medium for 24 h (Cat#05850, StemCell Technologies) containing 10 μM ROCK-inhibitor (Cat#Y0503, Sigma) and then switched to the differentiation medium: DMEM/F12 + GlutaMAX (Cat#31331, ThermoFisher), 17.5 mg/ml CellMaxx ...
The H9 hESCs were cultured on plates pre-coated with Matrigel (BD Biosciences, San Jose, CA) and maintained in the mTeSR™1 Complete Kit (Stem Cell Technologies, Vancouver, Canada) medium with daily medium change. When the cells reached 80% confluence, hESCs were detached from the plate wi...
and the pellet was resuspended into 1 ml of fresh complete medium. Typically the cells were plated at a density of 225 cells/100 mm dish. Substrate Preparation A. Fibronectin (FN) Substrate Tissue culture dishes were coated with human plasma fibronectin (New York Blood Center, New York, ...