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Primary cell-culture: Hepatocytes were isolated from liver of overnight fasted rat after subjecting it to two-stage collagenase perfusion with HEPES buffer (Seglen, 1976)

8 weeks: ~5 vials 12 weeks: ~7 vials 16 weeks: ~10 vials Insulin Syringes (U-100, 1 mL) Per injection: 1 syringe 8 weeks (once daily): ~56 syringes 16 weeks (once daily): ~112 syringes Bacteriostatic Water (10 mL bottles) Use ~3.0 mL per 10 mg vial for reconstitution

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