Pluripotent stem cells isolated from umbilical cord form embryonic like bodies in a mesenchymal layer culture.

Recently the matrix of umbilical cord began to use as an alternative source of stem cells additionally to the blood of umbilical cord. Umbilical cord has been used mainly for mesenchymal stem cell banking. The immunological characteristics of mesenchymal stem cells in combination with their ability to avoid rejection make them an attractive biological material for transplantations. In this study the isolation of small in size pluripotent stem cells from umbilical cord expressing early transcription factors with characteristics that resemble to embryonic stem cells is investigated. Pluripotent stem cells were isolated from human umbilical cords, by a new strategy method based on unique characteristics such as the small size and the positivity on early transcription factors OCT and Nanog. An enriched population of CXCR4(+) OCT(+) Nanog(+) CD45(-) small stem cells from the cord was isolated. This fraction was able to create alkaline phosphatase positive like spheres forms in a mesenchymal layer with multilineage differentiation capacity. Our results were assessed by RT PCR and electophoresis for the pluripotent genes. These data suggest that umbilical cord provides an attractive source not only of mesenchymal stem cells but moreover of pluripotent stem cells. The method described herein should be applied in the field of stem cell banking in addition to the classical umbilical cord harvesting method. Isolation of a population of cells with pluripotent characteristics from umbilical cord. Adoption of a second centrifugation step for the pluripotent stem isolation. Increasing the value of the cord and explaining the pluripotency. This work will enhance the value of umbilical cord harvesting.

A gene therapy induced emphysema model and the protective role of stem cells.

BACKGROUND: Chronic obstructive pulmonary disease presents with two different phenotypes: chronic bronchitis and emphysema with parenchymal destruction. Decreased expression of vascular endothelial growth factor and increased endothelial cell apoptosis are considered major factors for emphysema. Stem cells have the ability of vascular regeneration and function as a repair mechanism for the damaged endothelial cells. Currently, minimally invasive interventional procedures such as placement of valves, bio-foam or coils are performed in order to improve the disturbed mechanical function in emphysema patients. However, these procedures cannot restore functional lung tissue. Additionally stem cell instillation into the parenchyma has been used in clinical studies aiming to improve overall respiratory function and quality of life. METHODS: In our current experiment we induced emphysema with a DDMC non-viral vector in BALBC mice and simultaneously instilled stem cells testing the hyposthesis that they might have a protective role against the development of emphysema. The mice were divided into four groups: a) control, b) 50.000 cells, c) 75.000 and d) 100.000 cells. RESULTS: Lung pathological findings revealed that all treatment groups had less damage compared to the control group. Additionally, we observed that emphysema lesions were less around vessels in an area of 10 µm. CONCLUSIONS: Our findings indicate that stem cell instillation can have a regenerative role if applied upon a tissue scaffold with vessel around. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_195.

Rapidly progressing bilateral cataracts in a patient with beta thalassemia and pellagra.

Soon after the diagnosis of pellagra in a 20-year-old patient with beta thalassemia, bilateral intumescent cataracts rapidly developed. We believe the patient's crystalline lenses were at an increased oxidative state due to iron overload from the thalassemia. Depletion of the lens epithelial cells of an important antioxidative agent (glutathione) as a result of niacin (vitamin B3) deficiency due to pellagra reduced the antioxidative capacity of the lenses. The oxidative damage led to rapid development of cataracts.

Prebypass histological and ultrastructural evaluation of the long saphenous vein as a predictor of early graft failure.

BACKGROUND: Twenty percent of the long saphenous vein (LSV) grafts that are employed as coronary bypass conduits occlude during the first year after the operation. The aim of this study was to evaluate the morphological parameters of the LSV grafts before implantation as predictors for the early occlusion of the grafts. METHODS: Forty-two samples of LSV grafts were examined via light, transmission electron, and scanning electron microscopy and evaluated clinically and by angiography at 6 months and 2 years after the operation. Morphological parameters were statistically analyzed and examined for their significance on the viability of the vein grafts. RESULTS: Six (14.28%) of the examined grafts occluded within the first 6 months after the operation, and 11 grafts (26.19%) occluded within the first 2 years. The grafts that occluded at 6 months were characterized by thick intima (mean value, 206+/-32.29 vs. 67.44+/-10.17 in the group functioning normally and 98.42+/-34 in the group occluded within 2 years), low endothelial coverage (22.7+/-4.04 vs. 64.61+/-2.89 and 26.06+/-1.78 in the corresponding groups), and narrow lumen (46.73+/-9.69 vs. 527.18+/-45.78 and 204.26+/-16.5 in the corresponding groups). The presence of foam cells, edema, calcification, neovascularization, and thrombus in the lumen of the veins is frequently observed in the wall of the occluded vein grafts, whereas fibrosis does not seem to be related. CONCLUSIONS: LSV grafts with low endothelial cell coverage, stenosis of the lumen, and thick walls are at an increased risk of developing intrawall lesions that lead to early graft failure.

Laminin-1 is phosphorylated by ecto-protein kinases of monocytes.

Monocytes encounter basement membranes and interact with laminins while crossing the vascular barrier. It is known that these cells possess ecto-protein kinase activity on their surface. Several proteins of the extracellular matrix can be phosphorylated by ectokinases. Therefore, it has been hypothesized that monocyte ectokinases could phosphorylate laminins and influence their biological properties. In order to test the above hypothesis, we used intact human monocytes and adenosine triphosphate labeled with radioactive phosphate at the third phosphate ([gamma-32P]-ATP) to phosphorylate laminin-1. Autoradiography after sodium dodecyl sulphate polyacrylamyde gel electrophoresis (SDS-PAGE) electrophoresis indicated phosphorylation of laminin-1 on the beta and/or gamma chains. After phosphorylation, phosphoserine could be detected on Western blots by a specific monoclonal antibody. Phosphorylation was not detected when monocytes were pre-treated with trypsin and was inhibited by a specific ecto-protein kinase inhibitor (K252b). Laminin phosphorylation was also inhibited by heparin, a known inhibitor of casein kinase II and by pretreatment of monocytes by a monoclonal anti-casein kinase II antibody. Heparin binding, cell attachment and proliferation, and monocyte migration were enhanced on the phosphorylated laminin-1 as compared to the non-phosphorylated controls. These data indicate that laminin-1 can be phosphorylated by monocyte casein kinase II type ectokinase. This phosphorylation influences important functions of laminin and therefore could provide an additional means for the interaction of monocytes with basement membranes.