Reversal of hyperglycemia in mice by using human expandable insulin-producing cells differentiated from fetal liver progenitor cells

Abstract

Beta-cell replacement is considered to be the most promising approach for treatment of type 1 diabetes. Its application on a large scale is hindered by a shortage of cells for transplantation. Activation of insulin expression, storage, and regulated secretion in stem͞progen-itor cells offers novel ways to overcome this shortage. We explored whether fetal human progenitor liver cells (FH) could be induced to differentiate into insulin-producing cells after expression of the pancreatic duodenal homeobox 1 (Pdx1) gene, which is a key regulator of pancreatic development and insulin expression in beta cells. FH cells possess a considerable replication capacity, and this was further extended by introduction of the gene for the catalytic subunit of human telomerase. Immortalized FH cells expressing Pdx1 activated multiple beta-cell genes, produced and stored considerable amounts of insulin, and released insulin in a regulated manner in response to glucose. When transplanted into hyperglycemic immunodeficient mice, the cells restored and maintained euglycemia for prolonged periods. Quantitation of human C-peptide in the mouse serum confirmed that the glycemia was normalized by the transplanted human cells. This approach offers the potential of a novel source of cells for transplantation into patients with type 1 diabetes.

Blastema cells derived from New Zealand white rabbit’s pinna carry stemness properties as shown by differentiation into insulin producing, neural, and osteogenic lineages representing three embryonic germ layers

“…In this regard, we have previously reported that SLCs, obtained from the New Zealand white rabbit's pinnas, showed SC characteristics such as high proliferation ability and expression of pluripotency markers Oct4 and Sox2 (Mahmoudi et al 2011). In the present study, the capacity of SLCs to differentiate into cells derived It has been shown that serum-free medium (Zalzman et al 2003) and b-mercaptoethanol (Kojima et al 2004) promote the expression of Nestin, a pancreatic precursor marker, in SCs. Several studies have also indicated that high glucose concentration (25 mM) facilitates differentiation of SCs toward pancreatic (Sun et al 2007;Neshati et al 2010).…”

Adult rat liver cells transdifferentiated with lentiviral IPF1 vectors reverse diabetes in mice: an ex vivo gene therapy approach

“…Bearing in mind that the effect reached significance only 3 weeks after transplantation, it is possible that the in vivo environment allows these engineered cells to further mature or differentiate into fully functional cells that are capable of lowering blood glucose levels. In support of this, equine lentiviral-mediated expression of PDX1 turned fetal human progenitor liver cells into insulin-producing cells in about 3 weeks [30], while pancreatic non-endocrine cells transfected with PDX1 expressed insulin and other beta cell specific genes in vitro, but were only able to actually secrete insulin after being 6 PDX1-transduced hepatocytes (black symbols, n=7). Control diabetic mice were injected with equivalent amount of transduced hepatocytes with lentiviral vector but without PDX1 insert (white triangles, n=2).…”

“…Another limitation of adenoviral delivery of PDX1 is that it results in only about one copy per liver cell. Fetal human progenitor liver cells became insulin-producing cells in response to expression of PDX1 via a lentivirus (equine infectious anaemia virus-based vector) [30]. However, this method has not been tested using mature adult hepatocytes or a human lentiviral vector.…”

Hepatic targeting of transplanted liver sinusoidal endothelial cells in intact mice

“…17,18 Studies using hematopoietic, mesenchymal, and epithelial cells indicate that analysis of cell engraftment and function is necessary to establish the fate of transplanted cells. [19][20][21] This process involves cell targeting in specific organs (e.g., to establish suitable routes of cell administration or to assess cell trafficking in individual organs).…”