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CHIR99021 was removed after 72 h and DE cells were cultured in RPMI/B27 + Insulin for 24 h ( For growth factor Definitive Endoderm (DE) differentiation, human iPSCs cultured in 6-well dishes to 60% confluence were washed sequentially with DPBS / and DMEM/FI2 medium (Thermo Fisher Scientific)

UK B12 treatment for B12 deficiency without neuro symptoms is 6 loading jabs over 2 weeks followed by a jab every 3 months for B12 deficiency with neuro symptoms present, it is a B12 loading jab every other day for as long as symptoms continue to get better (could mean loading jabs for weeks even months) then a jab every 2 months UK b12 documents BMJ B12 article bmj.com/content/349/bmj.g5226 BSH Cobalamin and Folate Guidelines b-s-h.org.uk/guidelines/gui