Bactec aeorobic and anaerobic culture vials

RBCs were tested for endotoxin and absence of microbial growth (aerobic, anaerobic and fungal cultures) before and after biotin labeling. Before and after processing, 0.5 mL of product was added to Bactec aeorobic and anaerobic culture vials (catalog numbers 442194 and 442265; BD) and one drop of product was added to an inhibitory mold agar slant with Gentamicin (L49; Hardy Diagnostics). Cultures were maintained for 14 days. Endotoxin testing was performed on the supernatant from the last wash after biotinylation using the Endosafe PTS system (catalog number PTS 2005F; Charles River, Fisher Scientific), according to the manufacturer’s instructions. To determine suitability for infusion, the endotoxin reading (in Endotoxin Units (EU)/mL) is multiplied by the product volume in milliliters. This result (total endotoxin units in the product) is then divided by the patient’s actual body weight to determine the endotoxin dose per kilogram. An endotoxin dose < 5.0 EU/kg was considered acceptable [9 ].

Product release criteria included a final product calculated RBC packed cell volume ≥ 25 mL and total endotoxin < 5EU/Kg recipient actual body weight ¹³ . Endotoxin testing was performed on the supernatant from the last wash after biotinylation using the Endosafe PTS system (Charles River, Fisher Scientific, Cat. No. PTS 2005F), according to the manufacturer’s instructions. RBC were also tested retrospectively for absence of microbial growth (aerobic, anaerobic and fungal) before and after biotin labeling. Before and after processing 0.5 mL of product was added to Bactec aeorobic and anaerobic culture vials (BD, Cat Nos. 442194 and 442265) and one drop of product was added to an Inhibitory mold agar slant with gentamicin (Hardy Diagnostics, L49). Cultures were maintained for 14 days.