Figure 1.

BALM Generation

(A) hiPSCs differentiated to DECs, mouse livers decellularized using DET method. DECs injected into multiple locations in individual lobes on day 6 of differentiation. Then livers were placed into bioreactor and cultured under perfusion. Hepatic specification stage: day 7 to day 11 followed by hepatic maturation. During the maturation stage, viral vectors were perfused through the vasculature. BALMs were kept in culture till day 27.

(B) Stand-alone bioreactor composed of a chamber with medium in a closed circuit with a roller pump. The medium is oxygenated by 5% CO₂/Air gas cylinder regulated by a flowmeter. The gas is humidified in a water bottle while an overflow bottle collects outflow humidity. A bubble trap before the entrance of the chamber eliminates air bubbles. The system is maintained at 37°C using a hot plate under the chamber and the bottles.

(C) Bioreactor macroscopic appearance.

(D) Representative graph of pH monitoring of BALM, black dotted bars mark media type changes, values are average of two experiments.

See also Figure S1.