Amount and Distribution of Collagen in Pancreatic Tissue of Different Species in the Perspective of Islet Isolation Procedures

1995 ◽  
Vol 4 (6) ◽  
pp. 609-614 ◽  
Author(s):  
Paul T.R. van Suylichem ◽  
Jan-Erik H.M. van Deijnen ◽  
Gerrit H.J. Wolters ◽  
Reinout van Schilfgaarde
Keyword(s):  
Colorimetric Method ◽  
Pancreatic Tissue ◽  
Collagen Content ◽  
Human Pancreas ◽  
Islet Isolation ◽  
Secondary Importance ◽  
Enzymatic Dissociation ◽  
Tissue Sections ◽  
Dog Pancreas ◽  
General Experience

Because collagen is the major target in the enzymatic dissociation of the pancreas for islet isolation, we determined the amount of collagen and its distribution in a comparative study comprising normal pancreata of rat, dog, man, young pig, and adult pig. Collagen content was determined using a colorimetric method and its distribution was assessed in tissue sections stained with Sirius red. The collagen content is relatively low in the rat and adult pig pancreas, and the amount of collagen is relatively low in the septa of the rat and dog pancreas. Not the amount of collagen in the septa but collagen in the rest of the pancreas, mainly located between the acini, seems to determine the dissociation of the pancreatic tissue. This can be exemplified by the higher islet yields obtained from the adult vs. the young pig pancreas; the latter contains a higher total amount of collagen but a similar, relatively high, amount of collagen in the septa. A high amount of collagen surrounding the islets seems to be of secondary importance in islet isolations, because yields of the same magnitude are obtained from the canine and human pancreas containing a relatively low vs. high amount of collagen around the islets but a similar total collagen content. The rat pancreas contains both a low total amount of collagen and a high amount of collagen around the islets; therefore, the general experience that islet isolation procedures are effective in rats can be readily understood.

scholarly journals Role of eosinophils in the initiation and progression of pancreatitis pathogenesis

2018 ◽  
Vol 314 (2) ◽  
pp. G211-G222 ◽  
Author(s):  
Murli Manohar ◽  
Alok K. Verma ◽  
Sathisha Upparahalli Venkateshaiah ◽  
Anil Mishra
Keyword(s):  
Chronic Pancreatitis ◽  
Mast Cells ◽  
Critical Role ◽  
Pancreatic Tissue ◽  
Experimental Models ◽  
Human Pancreas ◽  
Protein Levels ◽  
Tissue Sections ◽  
Tissue Eosinophilia

Eosinophilic pancreatitis (EP) is reported in humans; however, the etiology and role of eosinophils in EP pathogenesis are poorly understood and not well explored. Therefore, it is interesting to examine the role of eosinophils in the initiation and progression of pancreatitis pathogenesis. Accordingly, we performed anti-major basic protein immunostaining, chloroacetate esterase, and Masson’s trichrome analyses to detect eosinophils, mast cells, and collagen in the tissue sections of mouse and human pancreas. Induced eosinophils accumulation and degranulation were observed in the tissue sections of human pancreatitis, compared with no eosinophils in the normal pancreatic tissue sections. Similarly, we observed induced tissue eosinophilia along with mast cells and acinar cells atrophy in cerulein-induced mouse model of chronic pancreatitis. Additionally, qPCR and ELISA analyses detected induced transcript and protein levels of proinflammatory and profibrotic cytokines, chemokines like IL 5, IL-18, eotaxin-1, eotaxin-2, TGF-β1, collagen-1, collagen-3, fibronectin, and α-SMA in experimental pancreatitis. Mechanistically, we show that eosinophil-deficient GATA1 and endogenous IL-5-deficient mice were protected from the induction of proinflammatory and profibrotic cytokines, chemokines, tissue eosinophilia, and mast cells in a cerulein-induced murine model of pancreatitis. These human and experimental data indicate that eosinophil accumulation and degranulation may have a critical role in promoting pancreatitis pathogenesis including fibrosis. Taken together, eosinophil tissue accumulation needs appropriate attention to understand and restrict the progression of pancreatitis pathogenesis in humans.NEW & NOTEWORTHY The present study for the first time shows that eosinophils accumulate in the pancreas and promote disease pathogenesis, including fibrosis in earlier reported cerulein-induced experimental models of pancreatitis. Importantly, we show that GATA-1 and IL-5 deficiency protects mice form the induction of eosinophil active chemokines, and profibrotic cytokines, including accumulation of tissue collagen in an experimental model of pancreatitis. Additionally, we state that cerulein-induced chronic pancreatitis is independent of blood eosinophilia.

scholarly journals Evaluating the Effect of Serine Proteases on Collagenase Activity during Human Islet Isolation

2002 ◽  
Vol 11 (8) ◽  
pp. 821-826 ◽  
Author(s):  
Natisha L. Rose ◽  
Monica M. Palcic ◽  
Jonathan R. T. Lakey
Keyword(s):  
Serine Proteases ◽  
Islet Cells ◽  
Pancreatic Tissue ◽  
Human Islet ◽  
Activity Levels ◽  
Human Pancreas ◽  
Islet Isolation ◽  
Collagenase Activity ◽  
Collagenase Digestion ◽  
Endogenous Enzymes

Inconsistencies in human islet yields after collagenase digestion have been attributed to the activation of endogenous enzymes of the donor pancreas. It has been suggested that pancreatic serine proteases contribute to the proteolysis of collagenase. This study defined the effects of endogenous enzymes within the pancreas on pancreas dissociation during collagenase digestion. Levels of collagenase activity from samples taken throughout several steps in islet isolation procedures, both with and without the addition of the serine protease inhibitor Pefabloc, were determined by a spectrophotometric assay using N-[3-(2-furyl)acryloyl]-Leu-Gly-Pro-Ala as the substrate. Results clearly demonstrated that the level of collagenase activity remains stable throughout the isolation procedure despite differences in the donor factors from several cadaveric donor pancreases. This was further demonstrated by observing no difference in activity levels after incubating commercial collagenase preparations with serine proteases and analyzing by means of collagenase activity and SDS-PAGE. These data show that the presence of serine proteases does not affect the level of collagenase activity; however, they likely damage the islet cells upon prolonged digestion of the pancreatic tissue. Further efforts at examining exogenous and endogenous enzyme levels may result in the development of an enzyme cocktail that is both stable and effective for digesting the human pancreas while preserving islet function and viability.

scholarly journals Pancreatic-Polypeptide in the Human Pancreas: Expression and Quantitative Variation During Development and in Ductal Adenocarcinoma

2003 ◽  
Vol 46 (1) ◽  
pp. 9-14 ◽  
Author(s):  
Demetrio Tamiolakis ◽  
Constantine Simopoulos ◽  
Athanasia Kotini ◽  
Ioannis Venizelos ◽  
Theodoros Jivannakis ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Pancreatic Polypeptide ◽  
Pancreatic Tissue ◽  
Ductal Adenocarcinoma ◽  
Human Pancreas ◽  
Neoplastic Tissue ◽  
Mantle Zone ◽  
Tissue Sections ◽  
Ductal Pancreatic Adenocarcinoma ◽  
Significant Difference

Aim: To determine the immunoreactivity of pancreatic-polypeptide (PP) during the development of the human fetal pancreas and ductal pancreatic adenocarcinoma, given that, PP positive cells were demonstrated either into its embryonic anlage or into pancreatic cancer. Methods: Tissue sections from 15 pancreatic fetal specimens, and equal number of ductal adenocarcinoma specimens, were assessed. Results: The density of positive cells in the primitive exocrine ductal epithelium and endocrine epithelium was significantly higher than the relevant density in the neoplastic pancreatic tissue of mixed (ductal – endocrine) and pure ductal type (p1=0.001, p2<0.0005, p3 =0.046 and p4<0.0005 respectively). The above values were estimated during the 10th to 12th week. There was no significant difference in the density of positive cells in the mantle zone of the islets from the 13th to the 24th week, and the neoplastic tissue of mixed (p5=0.11) and pure ductal type (p6=0.23). Conclusion: The immunostaining for PP identifies a subgroup of pancreatic ductal adenocarcinomas with a neuroendocrine component, initially considered as pure ductal tumors, and mixed ductal and neuroendocrine tumors. This pattern of expression in neoplasms recapitulates the normal pattern during the embryonal development of the organ, raising the question of therapeutic efficacy of PP and analogues as potential adjuvant treatment of pancreatic cancer.

scholarly journals 1H MAS NMR of Donor Pancreatic Tissue is a Useful Predictor of Islet Viability Prior to Islet Isolation

2020 ◽  
Author(s):  
Lisa M.H. Tanguay ◽  
Carolyn M. Slupsky ◽  
Toshiaki Toshiaki ◽  
Bruce Lix ◽  
Brian D. Sykes ◽  
...  
Keyword(s):  
Nmr Spectra ◽  
Pancreatic Tissue ◽  
Mas Nmr ◽  
Human Pancreas ◽  
Islet Isolation ◽  
Tissue Samples ◽  
1H Mas Nmr ◽  
University Of Wisconsin ◽  
Clinical Islet Transplantation ◽  
Glucose Responsiveness

AbstractA significant limitation and cost to any clinical islet program is the related to processing human pancreas and not recovering significant numbers of viable islets for clinical transplantation. The development of an assay system that could be utilized and provide an index of cell and tissue viability before islet isolation would provide a major impact on the scientific aspects of organ preservation and a huge cost saving to any clinical islet transplantation program.Metabolomic analysis by 1H MAS NMR was used to assess samples of donor pancreatic tissue taken prior to islet isolation. A significant correlation was observed between the ratio of the combined integrals of the sugar (3.5-4.5 ppm) and choline (3.0-3.5 ppm) regions to the integrals of the CH3 (0.9 ppm) and CH2 (1.3 ppm) peaks of the 1H MAS NMR spectra of pancreatic tissue samples taken prior to islet isolation and the glucose responsiveness, a measure of islet viability, of the isolated islets (P<0.05). The effect of the two-layer (University of Wisconsin solution/perfluorochemical [UW/PFC]) cold-storage method, previously shown to restore ischemically damaged pancreases by increasing oxygenation, was also studied using 1H MAS NMR spectroscopy. PFC recovery of the donor pancreas also correlated with an increase in the combined integrals of the sugar and choline regions to the CH3 and CH2 peaks of the 1H MAS NMR spectra (P<0.05). In addition, significant differences in the integrals of the sugar region and CH2 peaks were observed between the pre- and post-PFC samples (P<0.05). These results support the notion that specific metabolites observed in 1H MAS NMR can be used as a means to assess reversible/irreversible tissue damage and offers a means to assess donor pancreatic tissue prior to islet isolation for transplantation.

scholarly journals Hypothermic oxygenated Machine Perfusion of the Human Pancreas for Clinical Islet Isolation: A Prospective Feasibility Study

2021 ◽  
Author(s):  
Jason B. Doppenberg ◽  
Marjolein Leemkuil ◽  
Marten A. Engelse ◽  
Christina Krikke ◽  
Eelco J.P. de Koning ◽  
...  
Keyword(s):  
Feasibility Study ◽  
Human Pancreas ◽  
Islet Isolation ◽  
Machine Perfusion

The analysis of surface saccharide profiles through fluorescein-labelled lectins in a rat pancreatic tissue with established metabolic syndrome model

2018 ◽  
Vol 44 (1) ◽  
pp. 98-104
Author(s):  
Yosun Mater ◽  
Sule Beyhan-Ozdas
Keyword(s):  
Metabolic Syndrome ◽  
Metabolic Diseases ◽  
Pancreatic Tissue ◽  
Control Group ◽  
High Fructose Diet ◽  
Tissue Sections ◽  
Membrane Surfaces ◽  
High Fructose ◽  
A Cell ◽  
And Control

Abstract“Glycans”, which are generally referred as oligosaccharides and polysaccharides, are structures that are present on all cellular surfaces with proteins and lipids being attached to their basic chain structures. Many studies in the field of glycobiology have identified the various and complicated biological roles of these glycans which make them perfect molecules to use in labelling and selecting body cells specifically. This study aims at analyzing the modifications in saccharide units of glycans on a cell membrane surfaces of the pancreatic tissue of rats to which normal and metabolic syndrome (MetS) are established. To this end, a MetS model was created through a high fructose diet in Spraque Dawley breed of rats and the pancreatic tissue sections of the group with MetS and control group animals were evaluated comparatively. The targeted saccharide units were examined with Fluorescent Microscope by using two different Fluorescein (FITC) labelled lectins, namely Maackia amurensis-1 lectin [FITC-(MAL-I)] and the Wheat Germ Agglutinin (FITC-WGA). It was observed that FITC-MAL-1-labelled Galβ4GlcNAc units did not change much due to high- fructose diet. On the other hand, more GlcNAc, Neu5Ac and β-GlcNAc units which are labelled with FITC-WGA lectin increase in numbers in pancreatic sections of high fructose diet, compared to control group. Thus, a rapid and specific labelling method, which can identify surface saccharide sequences specifically, was developed. The method can be used in early diagnosis and/or treatment for metabolic diseases.

scholarly journals Mercury in Pancreatic Cells of People with and without Pancreatic Cancer

2020 ◽  
Vol 17 (23) ◽  
pp. 8990
Author(s):  
Roger Pamphlett ◽  
Andrew J. Colebatch ◽  
Philip A. Doble ◽  
David P. Bishop
Keyword(s):  
Pancreatic Cancer ◽  
Toxic Metals ◽  
Inductively Coupled Plasma ◽  
Islet Cells ◽  
Inorganic Mercury ◽  
Pancreatic Tissue ◽  
Human Pancreas ◽  
Inductively Coupled ◽  
Pancreatic Cells ◽  
Cadmium Lead

Toxic metals have been implicated in the pathogenesis of pancreatic cancer. Human exposure to mercury is widespread, but it is not known how often mercury is present in the human pancreas and which cells might contain mercury. We therefore aimed to determine, in people with and without pancreatic cancer, the distribution and prevalence of mercury in pancreatic cells. Paraffin-embedded sections of normal pancreatic tissue were obtained from pancreatectomy samples of 45 people who had pancreatic adenocarcinoma, and from autopsy samples of 38 people without pancreatic cancer. Mercury was identified using two methods of elemental bio-imaging: (1) With autometallography, inorganic mercury was seen in islet cells in 14 of 30 males (47%) with pancreatic cancer compared to two of 17 males (12%) without pancreatic cancer (p = 0.024), and in 10 of 15 females (67%) with pancreatic cancer compared to four of 21 females (19%) without pancreatic cancer (p = 0.006). Autometallographic mercury was present in acinar cells in 24% and in periductal cells in 11% of people with pancreatic cancer, but not in those without pancreatic cancer. (2) Laser ablation-inductively coupled plasma-mass spectrometry confirmed the presence of mercury in islets that stained with autometallography and detected cadmium, lead, chromium, iron, nickel and aluminium in some samples. In conclusion, the genotoxic metal mercury is found in normal pancreatic cells in more people with, than without, pancreatic cancer. These findings support the hypothesis that toxic metals such as mercury contribute to the pathogenesis of pancreatic cancer.

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