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Mouse livers were first perfused with Krebs-Ringer buffer containing 100 μM EGTA (MilliporeSigma) to chelate calcium, followed by enzymatic digestion using 60 mg of type I collagenase (Thermo Fisher Scientific) dissolved in 50 mL of DMEM/F12

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Where clinical suspicion remains high despite a normal serum B12, additional markers may be requested, including: Holotranscobalamin (active B12) — a more specific marker of B12 status, available in some UK laboratories as an alternative or confirmatory test [11] [12] Methylmalonic acid (MMA) — elevated in functional B12 deficiency