title: Reversal of diabetes following transplantation of an insulin-secreting human liver cell line: Melligen cells.
abstract: As an alternative to the transplantation of islets, a human liver cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial liver cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins cells with human islet glucokinase, the resultant Melligen cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins cells. When transplanted into diabetic immunoincompetent mice, Melligen cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both cell lines expressed a range of β-cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen cells in the presence of known β-cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.
authors.affiliation: School of Medical and Molecular Biosciences and Centre for Health Technologies, University of Technology Sydney , Sydney, Australia.
authors.forename: Janet
authors.initials: J
authors.lastname: Lawandi
authors.affiliation: School of Medical and Molecular Biosciences and Centre for Health Technologies, University of Technology Sydney , Sydney, Australia.
authors.forename: Chang
authors.initials: C
authors.lastname: Tao
authors.affiliation: School of Medical and Molecular Biosciences and Centre for Health Technologies, University of Technology Sydney , Sydney, Australia.
authors.forename: Binhai
authors.initials: B
authors.lastname: Ren
authors.affiliation: Department of Endocrinology, University of Sydney , Sydney, Australia.
authors.forename: Paul
authors.initials: P
authors.lastname: Williams
authors.affiliation: School of Medical and Molecular Biosciences and Centre for Health Technologies, University of Technology Sydney , Sydney, Australia.
authors.forename: Dora
authors.initials: D
authors.lastname: Ling
authors.affiliation: School of Medical Sciences (Anatomy & Histology) and Bosch Institute, University of Sydney , Sydney, Australia.
authors.forename: M Anne
authors.initials: MA
authors.lastname: Swan
authors.affiliation: School of Medical and Molecular Biosciences and Centre for Health Technologies, University of Technology Sydney , Sydney, Australia.
authors.forename: Najah T
authors.initials: NT
authors.lastname: Nassif
authors.affiliation: School of the Environment, University of Technology Sydney , Sydney, Australia.
authors.forename: Fraser R
authors.initials: FR
authors.lastname: Torpy
authors.affiliation: School of Medical and Molecular Biosciences and Centre for Health Technologies, University of Technology Sydney , Sydney, Australia.
authors.forename: Bronwyn A
authors.initials: BA
authors.lastname: O'Brien
authors.affiliation: School of Medical and Molecular Biosciences and Centre for Health Technologies, University of Technology Sydney , Sydney, Australia.
authors.forename: Ann M
authors.initials: AM
authors.lastname: Simpson