dithizone and Diabetes-Mellitus

Islet cell transplantation is an emerging therapy in the treatment of diabetes mellitus. Differentiation of islet cells from mesenchymal stem cells (MSCs) is a potential solution to the challenge of insufficient donor sources. This study investigated whether human umbilical cord-derived MSCs could effectively differentiate into insulin-producing cells (IPCs) and evaluated the therapeutic efficacy of IPCs in treating diabetes.. IPCs were induced from MSCs by a two-step protocol. IPC expression products were evaluated by western blot and real-time PCR. IPC insulin secretion was evaluated by ELISA. The viability of IPCs was measured by FDA/PI and dithizone staining. The non-human primate tree shrew was used as a diabetes model. After a single STZ induction into a diabetes model, a single intraportal transplantation of IPCs, MSCs, or normal saline was performed (n = 6 per group). Blood glucose was monitored for 3 weeks, then the animals were euthanized and the distribution of IPCs in the liver was examined pathologically.. After about 3 weeks of in vitro induction, IPCs formed microspheres of 100-200 μm, with >95% viable cells that were dithizone stain positive. IPCs expressed islet-related genes and proteins and secreted high levels of insulin whether stimulated by low or high levels of glucose. After transplantation of IPCs into diabetic tree shrews, blood glucose levels decreased rapidly to near normal and were significantly lower than the MSC or saline groups for 3 weeks thereafter.. We present the novel discovery that IPCs derived from human umbilical cord MSCs exert a therapeutic effect in a non-human primate model of diabetes. This study provides a preliminary experimental basis for the use of autologous MSC-derived IPCs in the treatment of human diabetes.

Topics: Animals; Blood Glucose; Diabetes Mellitus; Dithizone; Humans; Insulin; Primates

Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages in vitro. We have recently identified the emergence of cellular clusters within differentiated ES cell cultures by staining with dithizone (DTZ). DTZ is a zinc-chelating agent known to selectively stain pancreatic beta cells because of their high zinc content. The aim of the present study was to investigate the characteristics of DTZ-stained cellular clusters originating from ES cells.. Embryoid bodies (EBs), formed by a 5-day hanging drop culture of ES cells, were allowed to form outgrowths in the culture. The outgrowths were incubated in DTZ solution (final concentration, 100 microg/ml ) for 15 minutes before being examined microscopically. The gene expression of endocrine pancreatic markers was also analyzed by reverse transcriptase-polymerase chain reaction. In addition, insulin production was examined immunohistochemically, and its secretion was examined using enzyme-linked immunosorbent assay.. DTZ-stained cellular clusters appeared after approximately 16 days in the EB culture and became more apparent by day 23. They were found to be immunoreactive to insulin and expressed pancreatic-duodenal homeobox 1 (PDX1), proinsulin 1, proinsulin 2, glucagon, pancreatic polypeptide, glucose transporter-2 (GLUT2), and islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) mRNA. They were also able to secrete detectable amounts of insulin.. ES cell-derived DTZ-positive cellular clusters possess characteristics of the endocrine pancreas, including insulin secretion. Further, DTZ staining is a useful method for the identification of differentiated pancreatic islets developed from EBs in vitro.

Topics: Animals; Cell Culture Techniques; Cell Separation; Cells, Cultured; Chelating Agents; Diabetes Mellitus; Dithizone; Female; Glucose Transporter Type 2; Glucose-6-Phosphatase; Immunohistochemistry; Insulin; Insulin Secretion; Islets of Langerhans; Mice; Mice, Inbred Strains; Monosaccharide Transport Proteins; Proinsulin; Proteins; Stem Cell Transplantation; Totipotent Stem Cells; Zinc

In this study, diabetes was induced by intravenous injection of dithizone. In dithizonised diabetic animals, the levels of serum zinc, iron, and potassium were found to be higher than normal, while those of serum calcium and sodium were lower. Copper and magnesium levels were unchanged. After treatment with insulin, most of these serum levels approached the normal, except for serum potassium and magnesium.

Topics: Animals; Blood Glucose; Calcium; Copper; Diabetes Mellitus; Dithizone; Insulin; Iron; Minerals; Potassium; Rats; Sodium; Zinc

Topics: Animals; Diabetes Mellitus; Dithizone; Indicators and Reagents; Islets of Langerhans; Mice; Rabbits; Time Factors; Zinc

Topics: Animals; Blood Glucose; Cats; Cricetinae; Diabetes Mellitus; Dithizone; Dogs; Insulin; Insulin Secretion; Islets of Langerhans; Mice; Rabbits; Rats; Zinc

Topics: Adrenalectomy; Animals; Cell Count; Diabetes Mellitus; Dithizone; Epithelium; Islets of Langerhans; Pancreas; Rabbits

Topics: Animals; Diabetes Mellitus; Dithizone; Islets of Langerhans; Pathology; Physiology; Rabbits; Regeneration; Research

Topics: Animals; Diabetes Mellitus; Diabetes Mellitus, Experimental; Dithizone; Glycoproteins

Topics: Animals; Diabetes Mellitus; Diabetes Mellitus, Experimental; Dithizone; Indicators and Reagents; Oxyquinoline