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islet isolation

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https://www.readbyqxmd.com/read/27923539/the-roles-of-the-co-culture-of-mescs-with-pancreatic-islets-and-liver-stromal-cells-in-the-differentiation-of-definitive-endoderm-cells
#1
Hoveizi Elham, Fathi Fardin, Hashemitabar Mahmod
Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages in vitro. Investigating the roles of the co-culture of mouse embryonic stem cells (mESCs) with pancreatic islets (PL) and liver stromal cells (LSCs) in the differentiation of definitive endoderm (DE) cells was the purpose of this study. Here by using PL derived from adult mouse and LSCs derived from mouse fetal liver, we are trying to introduce a new protocol which is devoid of growth factors. We calculated mESCs indirectly for 7 days and then we analyzed the resulting cells regarding DE genes and protein expression using qRT-PCR and immunocytochemistry...
December 3, 2016: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/27920473/na-hco3-cotransporter-is-expressed-on-%C3%AE-and-%C3%AE-cells-during-rat-pancreatic-development
#2
Li-Hua Cao, Cheng-Cai Xia, Zhao-Chun Shi, Ning Wang, Zheng-Hua Gu, Li-Zhi Yu, Qi Wan, Wei De
AIM: To determine the expression and localization of the electrogenic Na(+)/HCO3(-) cotransporter (NBC1) in rat pancreas during development. METHODS: The rat pancreas from postnatal and embryos removed from the uterus of pregnant rats that had been sacrificed by CO2 asphyxiation were used. Rat pancreas from embryonic day (E) 15.5 and E18.5 rat embryos was isolated under a stereomicroscope. Rat pancreas from postnatal (P) days 0, 7, 14, 21 and adult was directly isolated by the unaided eye...
November 21, 2016: World Journal of Gastroenterology: WJG
https://www.readbyqxmd.com/read/27914538/adaptive-response-of-rat-pancreatic-%C3%AE-cells-to-insulin-resistance-induced-by-monocrotophos-biochemical-evidence
#3
Raju Nagaraju, Padmanabhan Sharda Rajini
Our previous findings clearly suggested the role of duration of exposure to monocrotophos (MCP) in the development of insulin resistance. Rats exposed chronically to MCP developed insulin resistance with hyperinsulinemia without overt diabetes. In continuation of this vital observation, we sought to delineate the biochemical mechanisms that mediate heightened pancreatic β-cell response in the wake of MCP-induced insulin resistance in rats. Adult rats were orally administered (0.9 and 1.8mg/kgb.w/d) MCP for 180days...
November 2016: Pesticide Biochemistry and Physiology
https://www.readbyqxmd.com/read/27909715/a-new-scaffold-containing-small-intestinal-submucosa-and-mesenchymal-stem-cells-improves-pancreatic-islet-function-and-survival-in%C3%A2-vitro-and-in%C3%A2-vivo
#4
Dan Wang, Xiaoming Ding, Wujun Xue, Jin Zheng, Xiaohui Tian, Yang Li, Xiaohong Wang, Huanjin Song, Hua Liu, Xiaohui Luo
It is unknown whether a scaffold containing both small intestinal submucosa (SIS) and mesenchymal stem cells (MSCs) for transplantation may improve pancreatic islet function and survival. In this study, we examined the effects of a SIS-MSC scaffold on islet function and survival in vitro and in vivo. MSCs and pancreatic islets were isolated from Sprague-Dawley rats, and SIS was isolated from Bamei pigs. The islets were apportioned among 3 experimental groups as follows: SIS-islets, SIS-MSC-islets and control-islets...
November 29, 2016: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/27892930/an-optimized-protocol-for-purification-of-functional-islets-of-langerhans
#5
Youakim Saliba, Jules-Joel Bakhos, Tarek Itani, Nassim Farès
Islets of Langerhans and β-cell isolation constitute routinely used cell models for diabetic research, and refining islet isolation protocols and cell quality assessment is a high priority. Numerous protocols have been published describing isolate of islets, but often rigorous and systematic assessment of their integrity is lacking. Herein, we propose a new protocol for optimal generation of islets. Pancreases from mice and rats were excised and digested using a low-activity collagenase solution and islets were then purified by a series of sedimentations and a Percoll gradient...
November 28, 2016: Laboratory Investigation; a Journal of Technical Methods and Pathology
https://www.readbyqxmd.com/read/27888197/islet-adaptations-in-fetal-sheep-persist-following-chronic-exposure-to-high-norepinephrine
#6
Xiaochuan Chen, Amy C Kelly, Dustin T Yates, Antoni R Macko, Ronald M Lynch, Sean W Limesand
Complications in pregnancy elevate fetal norepinephrine (NE) concentrations. Previous studies in NE-infused sheep fetuses revealed that sustained exposure to high NE resulted in lower expression of α2-adrenergic receptors in islets and increased insulin secretion responsiveness after acutely terminating the NE infusion. In this study, we determined if the compensatory increase in insulin secretion following chronic elevation of NE is independent of hyperglycemia in sheep fetuses and whether it is persistent in conjunction with islet desensitization to NE...
November 25, 2016: Journal of Endocrinology
https://www.readbyqxmd.com/read/27870582/markers-of-islet-endothelial-dysfunction-occur-in-male-db-db-mice-and-may-contribute-to-reduced-insulin-release
#7
Meghan F Hogan, Amy W Liu, Michael J Peters, Joshua R Willard, Zaheen Rabbani, Erik C Bartholomew, Adam Ottley, Rebecca L Hull
Islet endothelial cells produce paracrine factors that support β-cell function and growth. Endothelial dysfunction underlies diabetic microvascular complications; thus we hypothesized that in diabetes, islet endothelial cells become dysfunctional and this may contribute to β-cell secretory dysfunction. Islets/islet endothelial cells were isolated from diabetic db/db mice, treated with or without the glucose-lowering agent phlorizin, or C57BL/6J mice fed a high fat diet for 18 weeks (and appropriate controls)...
November 21, 2016: Endocrinology
https://www.readbyqxmd.com/read/27862363/a-comprehensive-microbiological-safety-approach-for-agarose-encapsulated-porcine-islets-intended-for-clinical-trials
#8
Lawrence S Gazda, James Collins, Archie Lovatt, Robert W Holdcraft, Merribeth J Morin, Daniel Galbraith, Melanie Graham, Melissa A Laramore, Christine Maclean, John Black, Euan W Milne, Douglas G Marthaler, Horatiu V Vinerean, Michelle M Michalak, Deborah Hoffer, Steven Richter, Richard D Hall, Barry H Smith
BACKGROUND: The use of porcine islets to replace insulin-producing islet β-cells, destroyed during the diabetogenic disease process, presents distinct challenges if this option is to become a therapeutic reality for the treatment of type 1 diabetes. These challenges include a thorough evaluation of the microbiological safety of the islets. In this study, we describe a robust porcine islet-screening program that provides a high level of confidence in the microbiological safety of porcine islets suitable for clinical trials...
November 11, 2016: Xenotransplantation
https://www.readbyqxmd.com/read/27862341/cost-and-clinical-outcome-of-islet-transplantation-in-norway-2010-2015
#9
Simen W Schive, Aksel Foss, Afaf Sahraoui, Kristine Kloster-Jensen, Geir Hafsahl, Gunnar Kvalheim, Torbjørn Lundgren, Bengt von Zur-Mühlen, Marie Felldin, Ehab Rafael, Marko Lempinen, Olle Korsgren, Trond G Jenssen, Vinod Mishra, Hanne Scholz
Islet transplantation is a minimally invasive β-cell replacement strategy. Islet transplantation is a reimbursed treatment in Norway. Here we summarize the cost and clinical outcome of 31 islet transplantations performed at Oslo University Hospital (OUS) from Januray 2010 - June 2015. Patients were retrospectively divided into 3 groups. 13 patients received either one or two islet transplantation alone (ITA), while 5 patients received islet transplantation after previous solid organ transplantation. For the group receiving 2 ITA, Kaplan-Meier estimates show an insulin independence of 20% more than 4 years after their last transplantation...
November 16, 2016: Clinical Transplantation
https://www.readbyqxmd.com/read/27844077/fe-oxy-adducts-of-heme-a%C3%AE-and-heme-hiapp-complexes-intermediates-in-ros-generation
#10
Manas Seal, Soumya Mukherjee, Somdatta Ghosh Dey
Iron (Fe) is the most abundant transition metal ion in the human body and its role, in the form of heme, has been implicated in Alzheimer's disease (AD) and type 2 diabetes mellitus (T2Dm). Heme binds both amyloid beta (Aβ) and human islet amyloid polypeptide (hIAPP) to form heme-Aβ and heme-hIAPP complexes, respectively, and form reactive oxygen species (ROS) like H2O2, O2˙(-)etc., which are known to cause oxidative damage. However the intermediates involved during ROS formation have not yet been isolated...
November 15, 2016: Metallomics: Integrated Biometal Science
https://www.readbyqxmd.com/read/27843954/continuous-quadrupole-magnetic-separation-of-islets-during-digestion-improves-purified-porcine-islet-viability
#11
Bradley P Weegman, Venkata Sunil Kumar Sajja, Thomas M Suszynski, Michael D Rizzari, William E Scott Iii, Jennifer P Kitzmann, Kate R Mueller, Thomas R Hanley, David J Kennedy, Paul W Todd, Appakalai N Balamurugan, Bernhard J Hering, Klearchos K Papas
Islet transplantation (ITx) is an emerging and promising therapy for patients with uncontrolled type 1 diabetes. The islet isolation and purification processes require exposure to extended cold ischemia, warm-enzymatic digestion, mechanical agitation, and use of damaging chemicals for density gradient separation (DG), all of which reduce viable islet yield. In this paper, we describe initial proof-of-concept studies exploring quadrupole magnetic separation (QMS) of islets as an alternative to DG to reduce exposure to these harsh conditions...
2016: Journal of Diabetes Research
https://www.readbyqxmd.com/read/27842980/establishment-characterization-and-long-term-culture-of-human-endocrine-pancreas-derived-microvascular-endothelial-cells
#12
Valeria Sordi, Anna Ferri, Valentina Ceserani, Emilio Ciusani, Erica Dugnani, Silvia Pellegrini, Rita Nano, Lorenza Pecciarini, Augusto Pessina, Luisa Pascucci, Lorenzo Piemonti, Giulio Alessandri
BACKGROUND: In vitro primary cultures of microvascular endothelial cells from endocrine pancreas are difficult to obtain, but can be a very helpful tool for studies of islet biology, transplantation and regenerative medicine. METHODS: We applied a protocol recently described for the isolation and culture of brain microvascular endothelial cells (EC) on human pancreatic islets. EC obtained were characterized in terms of morphological (light and transmission electron microscopy), phenotypical (by immunofluorescence and flow cytometry) and functional (cord formation assay and protein secretion by multiplex bead-based assay) characteristics...
November 11, 2016: Cytotherapy
https://www.readbyqxmd.com/read/27834384/clinical-pancreatic-islet-transplantation
#13
REVIEW
A M James Shapiro, Marta Pokrywczynska, Camillo Ricordi
Clinical pancreatic islet transplantation can be considered one of the safest and least invasive transplant procedures. Remarkable progress has occurred in both the technical aspects of islet cell processing and the outcomes of clinical islet transplantation. With >1,500 patients treated since 2000, this therapeutic strategy has moved from a curiosity to a realistic treatment option for selected patients with type 1 diabetes mellitus (that is, those with hypoglycaemia unawareness, severe hypoglycaemic episodes and glycaemic lability)...
November 11, 2016: Nature Reviews. Endocrinology
https://www.readbyqxmd.com/read/27832535/epigenetic-analysis-of-endocrine-cell-subtypes-from-human-pancreatic-islets
#14
Jia Zhang, Klaus H Kaestner
Epigenetic mechanisms have been proposed to contribute to multiple complex diseases, including diabetes mellitus. Dissecting the epigenomic landscape of normal and disease states has greatly expanded our understanding of the regulation of key players in disease pathogenesis and is thus opening doors to novel therapeutic avenues. The human pancreatic islet is a pivotal micro-organ that maintains global glucose homeostasis. The heterogeneity of the islet impedes meaningful in-depth epigenetic analysis using whole islet tissue, because important marks in one cell type are masked by signals from other cell types...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27830180/optimizing-porcine-islet-isolation-to-markedly-reduce-enzyme-consumption-without-sacrificing-islet-yield-or-function
#15
Robert W Holdcraft, Michael L Green, Andrew G Breite, Lisa Circle, Eric D Meyer, Hollie Adkins, Steven G Harbeck, Barry H Smith, Lawrence S Gazda
BACKGROUND: Human allogeneic islet transplantation for treatment of type 1 diabetes provides numerous clinical benefits, such as fewer episodes of hypoglycemic unawareness and tighter control of blood glucose levels. Availability of human pancreas for clinical and research use, however, is severely limited. Porcine pancreas offers an abundant source of tissue for optimization of islet isolation methodology and future clinical transplantation, thereby increasing patient access to this potentially lifesaving procedure...
July 2016: Transplantation Direct
https://www.readbyqxmd.com/read/27823612/islet-cell-transplantation-and-alternative-therapies
#16
REVIEW
Betul Hatipoglu
Even though type 2 diabetes rates plateaued, type 1 diabetes continues to increase. Pancreas transplantation is a treatment modality for patients who suffer hypoglycemic unawareness or complications from diabetes. Islet cell transplantation success rates have improved with modification and advances in isolation, transplantation, and new immunosuppression regimens. The new cell sources as well as delivery ways are explored and being tested in human trials.
December 2016: Endocrinology and Metabolism Clinics of North America
https://www.readbyqxmd.com/read/27820841/metformin-ameliorates-dysfunctional-traits-of-glibenclamide-and-glucose-induced-insulin-secretion-by-suppression-of-imposed-overactivity-of-the-islet-nitric-oxide-synthase-no-system
#17
Ingmar Lundquist, Israa Mohammed Al-Amily, Sandra Meidute Abaraviciene, Albert Salehi
Metformin lowers diabetic blood glucose primarily by reducing hepatic gluconeogenesis and increasing peripheral glucose uptake. However, possible effects by metformin on beta-cell function are incompletely understood. We speculated that metformin might positively influence insulin secretion through impacting the beta-cell nitric oxide synthase (NOS)-NO system, a negative modulator of glucose-stimulated insulin release. In short-time incubations with isolated murine islets either glibenclamide or high glucose augmented insulin release associated with increased NO production from both neural and inducible NOS...
2016: PloS One
https://www.readbyqxmd.com/read/27818940/critical-role-for-adenosine-receptor-a2a-in-%C3%AE-cell-proliferation
#18
Nadja Schulz, Ka-Cheuk Liu, Jérémie Charbord, Charlotte L Mattsson, Lingjie Tao, Dominika Tworus, Olov Andersson
OBJECTIVE: Pharmacological activation of adenosine signaling has been shown to increase β-cell proliferation and thereby β-cell regeneration in zebrafish and rodent models of diabetes. However, whether adenosine has an endogenous role in regulating β-cell proliferation is unknown. The objective of this study was to determine whether endogenous adenosine regulates β-cell proliferation-either in the basal state or states of increased demand for insulin-and to delineate the mechanisms involved...
November 2016: Molecular Metabolism
https://www.readbyqxmd.com/read/27816562/over-expression-of-insulin-response-element-binding-protein-1-ire-bp1-in-mouse-pancreatic-islets-increases-expression-of-rack1-and-tctpp-beta-cell-markers-of-high-glucose-sensitivity
#19
Betty C Villafuerte, Michelle T Barati, Madhavi J Rane, Susan Isaacs, Ming Li, Daniel W Wilkey, Michael L Merchant
BACKGROUND: A targeted analysis of the 50kDa C-terminal fragment of insulin-response element binding protein-1 (IRE-BP1) activation of target genes through the insulin receptor substrate receptor/PI-3 kinase/Akt pathway has been demonstrated for the insulin growth factor-1 receptor. The broader effects of 50kDa C-terminal IRE-BP1 fragment over-expression on protein abundance in pancreatic islet beta cells have not been determined. RESULTS: Liquid-chromatography coupled to tandem mass spectrometry (LC-MS/MS) analyses of replicate lysates of pancreatic islets isolated from background strain animals and transgenic animals, overexpressing IRE-BP1 in pancreatic islet beta cells, demonstrated statistically significant increases in the expression of proteins involved in protein synthesis, endoplasmic reticulum (ER) stress and scaffolding proteins important for protein kinase C signaling; some of which were confirmed by immunoblot analyses...
November 3, 2016: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27812538/proproliferative-and-antiapoptotic-action-of-exogenously-introduced-yap-in-pancreatic-%C3%AE-cells
#20
Ting Yuan, Sahar Rafizadeh, Zahra Azizi, Blaz Lupse, Kanaka Durga Devi Gorrepati, Sushil Awal, Jose Oberholzer, Kathrin Maedler, Amin Ardestani
Loss of functional pancreatic β cells is a hallmark of both type 1 and 2 diabetes. Identifying the pathways that promote β cell proliferation and/or block β cell apoptosis is a potential strategy for diabetes therapy. The transcriptional coactivator Yes-associated protein (YAP), a major downstream effector of the Hippo signaling pathway, is a key regulator of organ size and tissue homeostasis by modulating cell proliferation and apoptosis. YAP is not expressed in mature primary human and mouse β cells. We aimed to identify whether reexpression of a constitutively active form of YAP promotes β cell proliferation/survival...
November 3, 2016: JCI Insight
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