Differentiation of human skin-derived precursor cells into functional islet-like insulin-producing cell clusters

Abstract

Advances in cell-replacement strategies for diabetes have focused on renewable sources of glucose-responsive, insulin-producing cells (IPCs). One of the most proper alternatives is multipotent skin-derived precursors cells (SKPs), which can be differentiated into IPCs. In this study, we reported the isolation and expansion of human skin-derived precursors (hSKPs) followed by their differentiation into IPCs in vitro, through exposure to suitable differentiation factors. The gene expression of endocrine β cell markers was analyzed by reverse transcriptase-polymerase chain reaction. In addition, insulin production was examined immunocytochemically, and insulin and C-peptide secretion were examined using enzyme-linked immunosorbent assay. Dithizone-stained cellular clusters were observed after approximately 20 d. The clusters were found to be immunoreactive to insulin and expressed multiple genes related to pancreatic β cell development and function: insulin, Pdx-1, Islet-1, NeuroD, Glut-2, Pax-4, Ngn-3, and Nkx2.2, but not to other pancreas-specific hormones such as glucagon and somatostatin. Cellular clusters were also able to secrete detectable amounts of insulin and C-peptide in a glucose dose-dependent manner. These findings suggest that human SKPs can differentiate into functional IPCs. This may offer a safer cell source for future stem cell-based therapies.

Inflammation Alters the Secretome and Immunomodulatory Properties of Human Skin-Derived Precursor Cells

“…Human skin-derived precursors are a promising postnatal stem cell population for cellular therapy, as they exhibit multilineage differentiation capacity [5,7,8,16,19,22]. Furthermore, we previously showed that SKP are poorly immunogenic and exhibit favorable immunosuppressive properties [23].…”

“…SKP are of particular interest for regenerative medicine as they share several properties with embryonic neural crest-derived stem cells, a group of highly plastic cells with unique properties that reside within the developing embryo [2,12,13]. In particular, we and others have shown that SKP are able to generate (1) neuronal cells such as Schwann cells [7,14,15], catecholaminergic [16], dopaminergic [17] and enteric [18] neurons, (2) mesodermal cell types including adipocytes, chondrocytes, osteocytes [2] and vascular smooth muscle cells [19] and (3) endodermal progeny such as hepatic cells [5,9,20,21] and islet-like insulin-producing cells [22].…”

Adult muscle-derived stem cells engraft and differentiate into insulin-expressing cells in pancreatic islets of diabetic mice

“…Beta cells have been obtained from embryonic stems cells (ESC) of both mouse [8] and human origin [9] (reviewed in [10]), or from induced pluripotent stem cells (iPSC) produced through direct transcriptional reprogramming using stemness transcription factor cocktails (reviewed in [10–13]), or through reprogramming via a lineage conversion strategy from fibroblasts [14]. Although still controversial [15], a number of studies have described techniques for inducing the beta cell-specific reprogramming of fibroblasts of multiple origins into glucose-responsive beta-like cells [16–18]. However, long-term approaches involving differentiation of iPSC and ESC are compromised by the latent potential of residual tumor-promoting stem cells to develop into teratomas or cause tumorigenesis [19].…”

Stem Cells in the Treatment of InsulinDependent Diabetes Mellitus

“…Skin-derived precursors were converted in vitro into cells capable of glucose-dependent insulin and С-peptide secretion. The obtained cells expressed markers such as Pdx1, Nkx2.2, Pax4, NeuroD, and Isl1 found in mature β-cells [149].…”