title: Limited capacity of human adult islets expanded in vitro to redifferentiate into insulin-producing beta-cells.
abstract: Limited organ availability is an obstacle to the widespread use of islet transplantation in type 1 diabetic patients. To address this problem, many studies have explored methods for expanding functional human islets in vitro for diabetes cell therapy. We previously showed that islet cells replicate after monolayer formation under the influence of hepatocyte growth factor and selected extracellular matrices. However, under these conditions, senescence and loss of insulin expression occur after >15 doublings. In contrast, other groups have reported that islet cells expanded in monolayers for months progressed through a reversible epithelial-to-mesenchymal transition, and that on removal of serum from the cultures, islet-like structures producing insulin were formed (1). The aim of the current study was to compare the two methods for islet expansion using immunostaining, real-time quantitative PCR, and microarrays at the following time points: on arrival, after monolayer expansion, and after 1 week in serum-free media. At this time, cell aliquots were grafted into nude mice to study in vivo function. The two methods showed similar results in islet cell expansion. Attempts at cell differentiation after expansion by both methods failed to consistently recover a beta-cell phenotype. Redifferentiation of beta-cells after expansion is still a challenge in need of a solution.
authors.affiliation: Department of Pediatrics, USCD Whittier Institute, University of California-San Diego, 9894 Genesee Avenue, La Jolla, CA 92037-3495, USA.
authors.forename: Ayse G
authors.initials: AG
authors.lastname: Kayali
authors.forename: Luis E
authors.initials: LE
authors.lastname: Flores
authors.forename: Ana D
authors.initials: AD
authors.lastname: Lopez
authors.forename: Burak
authors.initials: B
authors.lastname: Kutlu
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Baetge
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Kitamura
authors.forename: Ergeng
authors.initials: E
authors.lastname: Hao
authors.forename: Gillian M
authors.initials: GM
authors.lastname: Beattie
authors.forename: Alberto
authors.initials: A
authors.lastname: Hayek
mesh_terms: D000818:Animals; D002096:C-Peptide / Q000097:blood / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D005786:Gene Expression Regulation; D005934:Glucagon / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000502:physiology; D007515:Islets of Langerhans / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude
chemical_list: D002096:C-Peptide; D007328:Insulin; D005934:Glucagon; D005947:Glucose