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The connective tissue and the gall bladder were removed, and the liver was minced into small pieces and forced gently through a 200-mm-gauge stainless steel mesh using a sterile syringe plunger and then suspended in 50 ml Roswell Park Memorial Institute (RPMI)-1640 medium containing GlutaMAX-1, 25 mM HEPES and 10% FCS (pH 7.4)

doi: 10.3390/cells9030561 148 WuRXiongJZhouTZhangZHuangZTianSet al

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Some protocols use a 'loading' phase (higher dose for 35 days) followed by maintenance dosing (lower dose long-term), based on the hypothesis that initial synaptogenesis requires stronger signaling, while maintaining new synapses requires less