title: Bioengineering the liver: scale-up and cool chain delivery of the liver cell biomass for clinical targeting in a bioartificial liver support system.
abstract: Acute liver failure has a high mortality unless patients receive a liver transplant; however, there are insufficient donor organs to meet the clinical need. The liver may rapidly recover from acute injury by hepatic cell regeneration given time. A bioartificial liver machine can provide temporary liver support to enable such regeneration to occur. We developed a bioartificial liver machine using human-derived liver cells encapsulated in alginate, cultured in a fluidized bed bioreactor to a level of function suitable for clinical use (performance competence). HepG2 cells were encapsulated in alginate using a JetCutter to produce ∼500 μm spherical beads containing cells at ∼1.75 million cells/mL beads. Within the beads, encapsulated cells proliferated to form compact cell spheroids (AELS) with good cell-to-cell contact and cell function, that were analyzed functionally and by gene expression at mRNA and protein levels. We established a methodology to enable a ∼34-fold increase in cell density within the AELS over 11-13 days, maintaining cell viability. Optimized nutrient and oxygen provision were numerically modeled and tested experimentally, achieving a cell density at harvest of >45 million cells/mL beads; >5×10(10) cells were produced in 1100 mL of beads. This process is scalable to human size ([0.7-1]×10(11)). A short-term storage protocol at ambient temperature was established, enabling transport from laboratory to bedside over 48 h, appropriate for clinical translation of a manufactured bioartificial liver machine.
authors.affiliation: Liver Group, UCL Institute of Liver & Digestive Health , London, United Kingdom .
authors.forename: Eloy
authors.initials: E
authors.lastname: Erro
authors.forename: James
authors.initials: J
authors.lastname: Bundy
authors.forename: Isobel
authors.initials: I
authors.lastname: Massie
authors.forename: Sherri-Ann
authors.initials: SA
authors.lastname: Chalmers
authors.forename: Aude
authors.initials: A
authors.lastname: Gautier
authors.forename: Spyridon
authors.initials: S
authors.lastname: Gerontas
authors.forename: Mike
authors.initials: M
authors.lastname: Hoare
authors.forename: Peter
authors.initials: P
authors.lastname: Sharratt
authors.forename: Sarah
authors.initials: S
authors.lastname: Choudhury
authors.forename: Marcin
authors.initials: M
authors.lastname: Lubowiecki
authors.forename: Ian
authors.initials: I
authors.lastname: Llewellyn
authors.forename: Cécile
authors.initials: C
authors.lastname: Legallais
authors.forename: Barry
authors.initials: B
authors.lastname: Fuller
authors.forename: Humphrey
authors.initials: H
authors.lastname: Hodgson
authors.forename: Clare
authors.initials: C
authors.lastname: Selden
keywords: HepG2 cells; alginate encapsulation; fluidized bed bioreactor