Comparison of biochemical and cytotoxic functions of hepatocytes from goat, pig and human fetuses

Vijayalakshmi, V.; Naseem, B.; Khan, Ansar A.; Capoor, A. K.; Habibullah, C. M.

doi:10.1111/j.1440-1746.2004.03402.x

Cited by 12 publications

(9 citation statements)

References 27 publications

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“…Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay [31]. Briefly, acinar cells (1X 10 5 ) were collected and incubated with PLP (5 mM) for 24 h and viability was measured by uptake of MTT reagent wavelength at 532 nm of all groups.…”

Section: Viability Assaymentioning

confidence: 99%

Beneficial Effects of Pyridoxal Phosphate on Hydrogen Peroxide Induced Stress on Isolated Pancreatic Acinar Cells

Ajumeera

Venkatesan

2016

Preprint

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Aims: To study the effects of pyridoxal phosphate (PLP) on oxidative stress in isolated pancreatic acinar cells. We have previously shown that PLP has cytoprotective and insulinotropic effects on mice islet cells in vivo and in vitro studies. Main methods: Acinar cells were isolated from three months old WNIN male rats and were cultured in vitro for a period of 24 h in CO2 incubator. Later the cells were divided into four groups as untreated (group 1), H2O2 treatment (group 2), PLP treatment (group 3) and PLP followed by H2O2 treatment (group 4). Cell viability was confirmed using MTT assays, oxidative stress levels were measured with ROS assay, change in different protein levels were recorded by flow cytometry. The acinar cells insulin secretion assay was performed with ELISA. The amylase protein expression was assessed using confocal microscopy. Key findings: The cell viability of acinar cell in group 1 was considered as 100%, while in group 2 it was reduced to 82% due to H2O2 effect, and in group 3 (99.8%) and group 4 (99.5%) were near to group 1 due to the cytoprotective effect of PLP. The ROS levels were increased by 1.47 folds in group 2, while PLP decreased to 1.02 fold in group 4, which was comparable with the changes in group 1. Beneficial effects of PLP were also observed from the increased expression levels of acinar cells are amylase -2.01, neurogenin-3-9.51, PDX-1-23.6 and insulin-13.5 in group 3 compared to group 1. The specificity of PLP's response was confirmed by amino oxy acetic acid (AOAA), a specific PLP inhibitor. The increased amylase protein localization with PLP was confirmed by confocal microscopy. Insulin secretion efficiency of acinar cells was observed to be 6.13 folds higher at basal and 24.63 fold higher at stimulated levels in group3 compared to group1. Significance: Our results advocate the antioxidant and cytoprotective effects of PLP on the pancreatic acinar cell along with increased pancreatic marker expressions of amylase,PDX-1, neurogenin-3 and insulin proteins.

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Section: Viability Assaymentioning

confidence: 99%

Beneficial Effects of Pyridoxal Phosphate on Hydrogen Peroxide Induced Stress on Isolated Pancreatic Acinar Cells

Ajumeera

Venkatesan

2016

Preprint

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“…15 In another set of experiments, islets (approximately 250) were pretreated with 2 mmol/L STZ and after 24h, PLP (Sigma) at a concentration of 1 -10 mmol/L was added and viability was assessed. 14 The primary islet cell cultures that were maintained in the media containing RPMI with 10% FCS were treated as controls.…”

Section: In Vitro Studiesmentioning

confidence: 99%

Pyridoxal 5′ phosphate protects islets against streptozotocin-induced beta-cell dysfunction – in vitro and in vivo

Kiran

Dorisetty

Umrani

et al. 2011

Exp Biol Med (Maywood)

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Administration of pyridoxal 5' phosphate (PLP) has demonstrated beneficial effects in the management of diabetes, albeit the mechanism(s) are not clearly understood. The present study addressed the islet-cell function(s) in streptozotocin (STZ)-induced diabetic mice both in vitro and in vivo. Primary islet cells primed with or without PLP (5 mmol/L) were treated with STZ (2 mmol/L) and were measured for cell viability, insulin secretion, free radicals and mRNA of Insulin and Pdx1. The specificity of PLP's response on insulin secretion was assessed with amino oxy acetic acid (AOAA)-PLP inhibitor. In vivo, the STZ (200 mg/kg b.w)-treated diabetic mice received 10 mmol/L PLP intraperitoneally a day before (PLP + STZ) or after (STZ + PLP) with three more doses once every 48 h. On 7, 14 and 21 d of STZ treatment, physiological parameters, islet morphology, insulin:glucagon, insulin:HSP104, and mRNA of Insulin, Glut2, Pdx1 and Reg1 were determined. In vitro, PLP protected islets against STZ-induced changes in viability, insulin secretion, prevented increase in free radical levels and normalized mRNA of Insulin and Pdx1. Further, AOAA inhibited PLP-induced insulin secretion in islets. In vivo, PLP treatment normalized STZ-induced changes in physiological parameters, circulating levels of PLP and insulin. Also, islet morphology, insulin:glucagon, insulin:HSP104 and mRNA levels of Insulin, Pdx1 and Glut2 were restored by 21 d. Although PLP treatment (pre- and post-STZ) prevented development of frank diabetes, STZ + PLP mice showed transient hyperglycemia, and increased mRNA for Reg1. The data suggest the cytoprotective vis-à-vis insulinotrophic effects of PLP against STZ-induced beta-cell dysfunction and underline its prophylactic use in the management of diabetes.

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“…Islet cell viability was assessed by Trypan Blue Dye Exclusion (TBE), the islet integrity was measured using Dithiazone (DTZ) (Sigma St. Louis, MO, USA) staining which helps one to differentiate islets from acinar cells (Banerjee and Bhonde 2003;Vijayalakshmi et al 2004). The number of islets in each group was determined by counting islets in triplicates (10% of the islet suspension) and multiplying the mean number of islets in these aliquots by 10.…”

Section: Viability and Integritymentioning

confidence: 99%

Optimization of condition(s) towards establishment of primary islet cell cultures from WNIN/Ob mutant rat

et al. 2011

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WNIN/Ob, a mutant rat strain, developed at the National Center for Laboratory Animal Sciences (NCLAS) facility of National Institute of Nutrition (NIN), is a new animal model to study the metabolic syndrome. These animals have 47% fat in their body and isolation of islets from these animals were compounded due to the formation of amorphous viscous and jelly like material which reduced the islet yield. However, islets isolated from WNIN adult (C12 months) control rats gave a good islet recovery, under standard isolation procedures using collagenase digestion. In the present study we optimized culture conditions in WNIN/Ob rats to isolate islets with higher yield, and also established primary islet cell cultures from these mutant rats, retaining cellular integrity and functionality.

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Comparison of biochemical and cytotoxic functions of hepatocytes from goat, pig and human fetuses

Abstract: These observations are significant as they suggest that goat hepatocytes probably can be explored as a source for cell therapy in the treatment of acute liver failure.

Cited by 12 publications

References 27 publications

Beneficial Effects of Pyridoxal Phosphate on Hydrogen Peroxide Induced Stress on Isolated Pancreatic Acinar Cells

Beneficial Effects of Pyridoxal Phosphate on Hydrogen Peroxide Induced Stress on Isolated Pancreatic Acinar Cells

Pyridoxal 5′ phosphate protects islets against streptozotocin-induced beta-cell dysfunction – in vitro and in vivo

Optimization of condition(s) towards establishment of primary islet cell cultures from WNIN/Ob mutant rat

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