Generation and expansion of multipotent mesenchymal progenitor cells from cultured human pancreatic islets

Abstract

Cellular models and culture conditions for in vitro expansion of insulin-producing cells represent a key element to develop cell therapy for diabetes. Initial evidence that human b-cells could be expanded after undergoing a reversible epithelialmesenchymal transition has been recently negated by genetic lineage tracing studies in mice. Here, we report that culturing human pancreatic islets in the presence of serum resulted in the emergence of a population of nestin-positive cells. These proliferating cells were mainly C-peptide negative, although in the first week in culture, proliferating cells, insulin promoter factor-1 (Ipf-1) positive, were observed. Later passages of islet-derived cells were Ipf-1 negative and displayed a mesenchymal phenotype. These human pancreatic islet-derived mesenchymal (hPIDM) cells were expanded up to 10 14 cells and were able to differentiate toward adipocytes, osteocytes and chondrocytes, similarly to mesenchymal stem/precursor cells. Interestingly, however, under serum-free conditions, hPIDM cells lost the mesenchymal phenotype, formed islet-like clusters (ILCs) and were able to produce and secrete insulin. These data suggest that, although these cells are likely to result from preexisting mesenchymal cells rather than b-cells, hPIDM cells represent a valuable model for further developments toward future replacement therapy in diabetes. Cell Death and Differentiation (2007) 14, 1860-1871; doi:10.1038/sj.cdd.4402199; published online 6 July 2007Type 1 diabetes mellitus is a chronic disease resulting from the selective autoimmune destruction of pancreatic insulinproducing b-cells. Transplantation therapy represents a potential cure for type 1 diabetes mellitus, 1 but is limited by availability of human pancreatic tissue. For this reason, a great effort has been made to develop new methods for generating b-like cells in vitro, 2,3 despite of evidence that cultured b-cells have limited proliferative capacity and reduced insulin production. 4 Several attempts have been made to identify stem/progenitors cells within pancreatic tissue as a potential source for transplantable insulinproducing tissue. Unfortunately, the origin of new b-cells in adult pancreas is not known. Some in vivo studies suggested the presence of pancreatic progenitor cells within islets, 5 whereas others reported that new adult b-cells might rather originate from pre-existing b-cells. 6 Additional studies suggested that progenitor cells may reside within the pancreatic ductal epithelium 2-7 or in the acinar tissue. 8,9 Nevertheless, the exact nature and localization of adult pancreatic stem/ progenitor cells remains controversial [9][10][11][12][13][14][15][16] and their existence in vivo, at least in mice, has recently been questioned. 6 Recent evidence has shown that human embryonic stem cells are able to differentiate into insulin-producing cells in vitro, thus potentially leading to an unlimited source of cells for transplantation. 17 These two authors contributed equally to this work. 5 These two authors share...

In Vitro Proliferation of Cells Derived From Adult Human β-Cells Revealed By Cell-Lineage Tracing

“…Initially, it was suggested that these cells originated from ␤-cells through epithelial-to-mesenchymal transition (EMT) (7). Recent work has documented the expression of mesenchymal stem cell (MSC) markers on these cells, and their ability to differentiate into osteocytes and adipocytes (20,21); however, their cellular origin has not been rigorously established. Using the labeling system described here, it should be possible to determine whether these cells are generated from ␤-cells by EMT or represent MSCs originally present in the islets.…”

Constant expression of hexose-6-phosphate dehydrogenase during differentiation of human adipose-derived mesenchymal stem cells

“…After 21 days of adipogenic stimulation, cells were fixed in 4% PFA for 1 h and incubated with Oil Red-O to stain lipid vacuoles. Chondrogenic and osteogenic differentiations were induced as described elsewhere (Gallo et al 2007). …”

Activin Receptor-Like Kinase 5 Inhibition Reverses Impairment of Endothelial Cell Viability by Endogenous Islet Mesenchymal Stromal Cells

“…In particular, hemopoietic precursors from bone marrow and bone marrow-derived MSCs both express high levels of endoglin, with expression fluctuating according to cell activation status [22,36,37]. Our characterization of endoglin-positive, EC marker-negative cells isolated and expanded from dispersed human islets showed that they exhibited MSC-like properties and expressed a range of MSC marker genes [38]. Islet MSCs also expressed high levels of Id-1 when compared to ECs, consistent with the high Id-1 mRNA expression detected in cultured islets.…”