Monolayer cultures of pancreatic tissue. IV. Investigations of insulin secretion and the influence of heterologous insulin

1972 ◽  
Vol 9 (1) ◽  
pp. 577-594 ◽  
Author(s):  
Ingeborg Hilwig
1988 ◽  
Vol 118 (2) ◽  
pp. 173-NP ◽  
Author(s):  
M. Aoki ◽  
S. Kagawa ◽  
T. Yamamura ◽  
A. Matsuoka ◽  
J. Utsunomiya

ABSTRACT Techniques for the monolayer culture of pancreatic islet cells from adult rats and the responsiveness of B cells are described. Whole pancreatic tissue was enzymatically dispersed and then cultured for 30 days in tissue culture medium 199 containing 5·5 mmol glucose/l, with or without 1 mmol 2-deoxyglucose/l. In the absence of 2-deoxyglucose, the responsiveness of B cells diminished to almost zero by day 15 and islets degenerated. In contrast, addition of 2-deoxyglucose to the medium resulted in a selective degeneration of fibroblasts, yielding monolayers that consisted mostly of islet cells. In this stationary system in which monolayers of islet cells were maintained in medium with 2-deoxyglucose, insulin secretion from B cells on days 15 and 30 increased in a dose-dependent fashion in response to increasing concentrations of glucose, leucine and 2-ketoisocaproate. Similarly, when exposed to 16·7 mmol glucose/l, perifused B cells showed a biphasic pattern of insulin secretion on day 15. Addition of 10 μmol forskolin/l and 200 nmol 12-O-tetradecanoyl phorbol13-acetate/l remarkably enhanced this response. Likewise, the response to 10 mmol leucine/l or 10 mmol 2-ketoisocaproate/l was biphasic. These results suggest that these monolayer cultures retain the functional properties of the adult rat pancreas, and may be useful not only as a model for the in-vitro study of B cell function, but also for implantation. J. Endocr. (1988) 118, 173–178


1978 ◽  
Vol 235 (3) ◽  
pp. E266 ◽  
Author(s):  
S Inoue ◽  
G A Bray ◽  
Y S Mullen

The present experiments have tested the hypothesis that ventromedial hypothalamic (VMH) lesions enhance insulin secretion by neural mechanisms. Rats were made diabetic by injecting streptozotocin to destroy their own pancreatic beta-cells. Subsequently, transplants of fetal pancreatic tissue were placed under the renal capsule. VMH lesions were placed in rats whose diabetes was cured with transplants as well as sham-transplanted animals. The animals were followed for 4 wk. The lesioned rats with pancreatic transplants gained no more weight than the sham-operated controls. There was no significant rise in insulin in the transplanted rats after VMH lesioning, but the VMH lesioned rats with intact pancreatic tissue showed the expected rise in insulin. Food intake rose 71% in the VMH lesioned rats with intact beta-cells, but only 23% in the VMH lesioned rats with transplants. Hypertrophy of the pancreatic islets was also observed in the VMH lesioned rats with an intact pancreas, but was not found in the VMH lesioned rats with a transplanted pancreas. Thus, transplantation of pancreatic tissue beneath the renal capsule of diabetic rats prevented the characteristic hyperphagia, hyperinsulinemia, and obesity in VMH lesioned rats whose pancreas was free from intact innervation. The results support the hypothesis that neural mediation of the rise in insulin is the primary factor in the development of hypothalamic obesity.


2011 ◽  
Vol 63 (4) ◽  
pp. 1007-1013
Author(s):  
D.M. Nikolic ◽  
P.B. Djordjevic ◽  
Dj. Radak ◽  
Vesna Lackovic ◽  
Milos Bajcetic ◽  
...  

In this study we compared the effects of duration of cold ischemia (longer and shorter ischemia) on the yield, viability and preservation of the physiological function and insulin secretion of adult human pancreatic islets in short-term (seven days) culture. Based on the tested parameters, we established that there are no major differences between these two test groups and that the storage and transport of pancreatic tissue in physiological solution at 4oC gives quite satisfactory results.


2019 ◽  
Vol 47 ◽  
Author(s):  
Ignacio Netzahualcoyotl Barajas-López ◽  
Jesús Aurelio Medina-Flores ◽  
Gabriela Arnaud-Pérez ◽  
Marco Antonio Mendoza-Rivera ◽  
Yurixhi López-Garcia ◽  
...  

Background: The most frequent pancreatic tumours are derived from insulin-secreting β cells, commonly called insulinomas; these are characterised by high insulin secretion causing hypoglycaemia and clinical signs such as seizures, tremors, weakness, and polyphagia, among others. In dogs, this tumour represents <0.5% of neoplasias; the majority are solitary carcinoma masses and rarely adenomas. Insulin-secreting tumours are usually diagnosed in middle-aged or older dogs. There is no apparent sex predilection for the disease and it has been mainly reported in medium to large breeds. Independently of whether they are adenomas or carcinomas, dogs have the same disease-free time and survival time and the prognosis is poor.Case: An 8-year-old female Boxer was brought to the University Veterinary Hospital with a history of weakness, tremors, and generalised convulsions. Physical examination, CBC and urinalysis revealed no abnormalities. In the blood chemistry profile, hypoglycaemia was detected along with hyperinsulinemia. An abdominal ultrasound revealed the presence of two abnormal masses located in the pancreas. The insulin:glucose ratio was 59.8. Exploratory celiotomy was performed and two masses were located in the mesentery, adjacent to the left pancreatic lobe, and a third was in the pancreatic tissue of the same lobule. All masses were resected during the same surgery. Cytology of the masses coincided with apparently malignant insulinoma, however, the histopathological and immunohistochemically report indicated an insulin-secreting adenoma. The patient improved clinically and remained stable for approximately 545 days, after which seizures relapsed and a new treatment was not approved. The patient died 575 days after surgery. Discussion: In dogs, pancreatic islet cell tumours correspond to endocrinologically active neoplasm that secrete hormones and are associated with functional disorders (hyperinsulinemia) in relation to hypoglycemia. Hypoglycemia causes episodic signs that are generally observed for a few seconds to several minutes, because of regulatory compensatory mechanisms. In animals, rapid hypoglycemia activates the sympathetic nervous system via hypothalamic glucoreceptors, producing signs such as tachycardia, tremors, nervousness, irritability, and intense hunger. Other clinical signs are related to neuroglycopenia by a decrease in blood glucose, which stimulates the autonomic nervous system, causing hypothermia, lethargy, weakness, ataxia, collapse, muscular fasciculation, convulsions, and coma. The diagnosis of an insulin-secreting tumour requires confirmation of hypoglycemia with evidence of elevated insulin secretion and the identification of a pancreatic mass by ultrasonography or exploratory celiotomy. An insulin:glucose ratio >30 is indicative of insulinoma. Unfortunately, the specificity of the amended insulin:glucose ratio is poor. In the past, there has been confusion with respect to their biological nature, because, based on histological and electron microscopic evaluations, 60% of these neoplasms are carcinomas and 40% are adenomas. Nevertheless, these claims were not substantiated and most insulinomas are currently considered to be malignant (carcinomas). The objective of this paper was to present a rare case of a Boxer dog that had three β cell tumours of the pancreas, which produced hyperinsulinemia and hypoglycemia. Although this is not a malignant tumour, we demonstrated, as described in the literature, that regardless of the histopathological classification, insulin-producing tumours have a poor prognosis in dogs.


2021 ◽  
Vol 3 (1) ◽  
pp. 1-13
Author(s):  
Anasuya Ganguly ◽  

Diabetes is a group of diseases characterized by high levels of blood sugar for an extended period. Despite newer and effective therapy, current treatment is riddled with fundamental challenges. To overcome the adverse effects of existing drugs, regenerative medicine has emerged as an essential treatment, for which tissue engineering may serve as a foundation for the repair of pancreatic cells secreting insulin. Different polymeric scaffolds have been explored for pancreatic tissue engineering. In the current study, a continuation of our preceding work we attempt to test the role of previously synthesized agarose-chitosan coated silver nanocomposite scaffold (AG-CHNp) for the long-term growth of pancreatic cells. Pancreatic cells were isolated from BALB/c mice and were characterized by dithizone (DTZ) staining, real time polymerase chain reaction (RT-PCR), western blotting, and flow cytometry for characteristic pancreatic markers. The isolated population of cells was grown on scaffolds and its effectiveness towards insulin secretion was studied. The isolated population was found to be positive for glucagon, PDX-1 and Pax-4, while a 200-fold change transcript level of insulin was observed. The cells upon seeding on the scaffolds exhibited sustained growth and insulin secretion as confirmed by western blotting. Overall, the study demonstrates the suitability and application of AG-CHNp for pancreatic tissue engineering.


2015 ◽  
Vol 35 (5) ◽  
pp. 1892-1904 ◽  
Author(s):  
Dan-dan Yin ◽  
Er-bao Zhang ◽  
Liang-hui You ◽  
Ning Wang ◽  
Lin-tao Wang ◽  
...  

Background: Increasing evidence indicates that long noncoding RNAs (IncRNAs) perform specific biological functions in diverse processes. Recent studies have reported that IncRNAs may be involved in β cell function. The aim of this study was to characterize the role of IncRNA TUG1 in mouse pancreatic β cell functioning both in vitro and in vivo. Methods: qRT-PCR analyses were performed to detect the expression of lncRNA TUG1 in different tissues. RNAi, MTT, TUNEL and Annexin V-FITC assays and western blot, GSIS, ELISA and immunochemistry analyses were performed to detect the effect of lncRNA TUG1 on cell apoptosis and insulin secretion in vitro and in vivo. Results: lncRNA TUG1 was highly expressed in pancreatic tissue compared with other organ tissues, and expression was dynamically regulated by glucose in Nit-1 cells. Knockdown of lncRNA TUG1 expression resulted in an increased apoptosis ratio and decreased insulin secretion in β cells both in vitro and in vivo . Immunochemistry analyses suggested decreased relative islet area after treatment with lncRNA TUG1 siRNA. Conclusion: Downregulation of lncRNA TUG1 expression affected apoptosis and insulin secretion in pancreatic β cells in vitro and in vivo. lncRNA TUG1 may represent a factor that regulates the function of pancreatic β cells.


1987 ◽  
Vol 10 (6) ◽  
pp. 547-552 ◽  
Author(s):  
R. Oosterom ◽  
T. Verleun ◽  
P. Uitterlinden ◽  
H. A. Brunning ◽  
G. J. Bruining ◽  
...  

2007 ◽  
Vol 129 (6) ◽  
pp. 493-508 ◽  
Author(s):  
Tobias Rose ◽  
Suad Efendic ◽  
Marjan Rupnik

The Goto Kakizaki (GK) rat is a widely used animal model to study defective glucose-stimulated insulin release in type-2 diabetes (T2D). As in T2D patients, the expression of several proteins involved in Ca2+-dependent exocytosis of insulin-containing large dense-core vesicles is dysregulated in this model. So far, a defect in late steps of insulin secretion could not be demonstrated. To resolve this apparent contradiction, we studied Ca2+–secretion coupling of healthy and GK rat β cells in acute pancreatic tissue slices by assessing exocytosis with high time-resolution membrane capacitance measurements. We found that β cells of GK rats respond to glucose stimulation with a normal increase in the cytosolic Ca2+ concentration. During trains of depolarizing pulses, the secretory activity from GK rat β cells was defective in spite of upregulated cell size and doubled voltage-activated Ca2+ currents. In GK rat β cells, evoked Ca2+ entry was significantly less efficient in triggering release than in nondiabetic controls. This impairment was neither due to a decrease of functional vesicle pool sizes nor due to different kinetics of pool refilling. Strong stimulation with two successive trains of depolarizing pulses led to a prominent activity-dependent facilitation of release in GK rat β cells, whereas secretion in controls was unaffected. Broad-spectrum inhibition of PKC sensitized Ca2+-dependent exocytosis, whereas it prevented the activity-dependent facilitation in GK rat β cells. We conclude that a decrease in the sensitivity of the GK rat β-cell to depolarization-evoked Ca2+ influx is involved in defective glucose-stimulated insulin secretion. Furthermore, we discuss a role for constitutively increased activity of one or more PKC isoenzymes in diabetic rat β cells.


1971 ◽  
Vol 49 (6) ◽  
pp. 536-544 ◽  
Author(s):  
Guy R. Brisson ◽  
Willy J. Malaisse

BS-4231, a new sulfonylurea, stimulates insulin secretion by incubated pieces of rat pancreatic tissue. The minimal effective dose is between 3 and 30 mμg/ml. The insulinotropic effect of this agent is transient. It is more marked at intermediate (1.0 and 1.5 mg/ml) than either at high glucose concentrations (3.0 mg/ml) or in the absence of glucose. Experimental data obtained with various inhibitors of insulin secretion suggest that (i) epinephrine suppresses the stimulant action of BS-4231, as well as that of glucose; (ii) mannoheptulose modulates the insulinotropic action of BS-4231 indirectly by altering the effect of extracellular glucose; and (iii) diazoxide faiis to exert any effect upon insulin secretion in the presence of BS-4231. On the basis of these results, it is suggested that BS-4231 might accelerate within the beta-cell some step of glucose metabolism, which is rate-limiting when the insulin secretory process is not fully stimulated by extracellular glucose.


1994 ◽  
Vol 6 (8) ◽  
pp. 897-904 ◽  
Author(s):  
Marsha A. Black ◽  
Geoffrey A.R. Mealing ◽  
James F. Whitfield ◽  
Jean-Louis Schwartz ◽  
Nicole Bégin-heick

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