The Experts below are selected from a list of 294 Experts worldwide ranked by ideXlab platform
Xudong Zhao - One of the best experts on this subject based on the ideXlab platform.
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Primary Culture of Porcine Pancreatic Acinar Cells
Pancreas, 2002Co-Authors: Chengwei Tang, Xudong ZhaoAbstract:INTRODUCTION: The methodology of acinar cell Culture has become of Primary importance in the research of pancreatic physiology and pharmacology. AIM: To develop a method for Primary Culture of porcine pancreatic acinar cells. METHODOLOGY: Dispersed pancreatic acinar cells were made by RPMI-1640 medium containing collagenase III. After purification, the isolated acinar cells were Cultured in RPMI-1640 medium with 2.5% fetal bovine serum. The morphologic characteristics of acinar cells were described. (3)H-thymidine incorporation of acinar cells and activity of amylase or lipase were determined during the Culture. RESULTS: There were no remarkable morphologic changes in the pancreatic acinar cells during the 20-day Culture. The acini showed the tendency of gathering but did not attach to the walls of the Culture disks. Incorporation of (3)H-thymidine in acinar cells in the Primary Culture was well kept. The secretion of amylase or lipase from acini decreased with the time of Culture. CONCLUSIONS: In the Primary Culture of acinar cells from porcine pancreas developed in this study, the acinar cells retained normal morphology and ability of growth but not secretion of amylase or lipase. The method would be beneficial for further experiments on acini of porcine pancreas.
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Primary Culture of Porcine Pancreatic Acinar Cells
Pancreas, 2002Co-Authors: Chengwei Tang, Xudong Zhao, Jun HanAbstract:Objective To develop a method for the Primary Culture of porcine pancreatic acinar cells. Interventions Dispersed pancreatic acinar cells available utilizing RPMI-1640 medium containing collagenase III. After purification, the isolated acinar cells were Cultured in RPMI1640 medium with the addition of 2.5% fetal bovine serum. Main outcome measures The morphological characteristics of acinar cells were described. 3 H-thymidine incorporation of acinar cells and the activity of amylase or lipase were determined during the Culture process. Results There were no remarkable morphological changes in the pancreatic acinar cells during the 20 days ’ Culture. The acini showed a tendency to gather but did not attach to the walls of the Culture disks. A good 3 Hthymidine incorporation of acinar cells in the Primary Culture was maintained. The secretion of amylase or lipase from the acini decreased with the length of time of the Culture. Discussion The Primary Culture of acinar cells from a porcine pancreas which was carried out in this study maintained the normal morphology of the acinar cells and the ir ability to grow but not the ir secretion of amylase or lipase. The method would benefit by the further experiments on acini of porcine pancreas.
Chengwei Tang - One of the best experts on this subject based on the ideXlab platform.
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Primary Culture of Porcine Pancreatic Acinar Cells
Pancreas, 2002Co-Authors: Chengwei Tang, Xudong ZhaoAbstract:INTRODUCTION: The methodology of acinar cell Culture has become of Primary importance in the research of pancreatic physiology and pharmacology. AIM: To develop a method for Primary Culture of porcine pancreatic acinar cells. METHODOLOGY: Dispersed pancreatic acinar cells were made by RPMI-1640 medium containing collagenase III. After purification, the isolated acinar cells were Cultured in RPMI-1640 medium with 2.5% fetal bovine serum. The morphologic characteristics of acinar cells were described. (3)H-thymidine incorporation of acinar cells and activity of amylase or lipase were determined during the Culture. RESULTS: There were no remarkable morphologic changes in the pancreatic acinar cells during the 20-day Culture. The acini showed the tendency of gathering but did not attach to the walls of the Culture disks. Incorporation of (3)H-thymidine in acinar cells in the Primary Culture was well kept. The secretion of amylase or lipase from acini decreased with the time of Culture. CONCLUSIONS: In the Primary Culture of acinar cells from porcine pancreas developed in this study, the acinar cells retained normal morphology and ability of growth but not secretion of amylase or lipase. The method would be beneficial for further experiments on acini of porcine pancreas.
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Primary Culture of Porcine Pancreatic Acinar Cells
Pancreas, 2002Co-Authors: Chengwei Tang, Xudong Zhao, Jun HanAbstract:Objective To develop a method for the Primary Culture of porcine pancreatic acinar cells. Interventions Dispersed pancreatic acinar cells available utilizing RPMI-1640 medium containing collagenase III. After purification, the isolated acinar cells were Cultured in RPMI1640 medium with the addition of 2.5% fetal bovine serum. Main outcome measures The morphological characteristics of acinar cells were described. 3 H-thymidine incorporation of acinar cells and the activity of amylase or lipase were determined during the Culture process. Results There were no remarkable morphological changes in the pancreatic acinar cells during the 20 days ’ Culture. The acini showed a tendency to gather but did not attach to the walls of the Culture disks. A good 3 Hthymidine incorporation of acinar cells in the Primary Culture was maintained. The secretion of amylase or lipase from the acini decreased with the length of time of the Culture. Discussion The Primary Culture of acinar cells from a porcine pancreas which was carried out in this study maintained the normal morphology of the acinar cells and the ir ability to grow but not the ir secretion of amylase or lipase. The method would benefit by the further experiments on acini of porcine pancreas.
V. Storch - One of the best experts on this subject based on the ideXlab platform.
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Senescence of hepatocytes isolated from rainbow trout (Oncorhynchus mykiss) in Primary Culture
Protoplasma, 1992Co-Authors: T. Braunbeck, V. StorchAbstract:In order to evaluate the suitability of isolated fish hepatocytes for toxicological studies, hepatocytes were isolated from rainbow trout ( Oncorhynchus mykiss ) by liver collagenase perfusion. Isolated hepatocytes were investigated for seven days in Primary Culture on uncoated petri dishes using light and electron microscopy. Viability of isolated hepatocytes, as estimated from trypan blue exclusion, declined from >90% at the beginning of the incubation to ⩽80% after eight days of Primary Culture. Survival of hepatocytes was best at an incubation temperature of 14°C, and addition of fetal calf serum failed to improve cell performance. Freshly isolated hepatocytes appeared as solitary spherical cells with numerous microvilli at the outer surface. Except for a 30% reduction in cell size due to stress-induced glycogen reduction, the ultrastructure of freshly isolated hepatocytes closely resembled that of rainbow trout hepatocytes in vivo, which is characterized by distinct cytoplasmic segregation into a perinuclear portion containing rough endoplasmic reticulum (RER) arranged in extensive stacks, mitochondria, peroxisomes and the peribiliary complex (dictyosomes, lysosomes), and large peripheral glycogen fields occasionally interspersed with small lipid inclusions. Within one day of Culture, about 60–80% of the isolated hepatocytes sedimented to form a monolayer attached to the Culture dishes, whereas up to 20% remained in suspension forming hepatocyte aggregates. Cell adhesion was weak, and during prolonged Culture increasing amounts of cell detached, whereas the floating cell accumulations grew to aggregates of more than 100 cells. Cell viability and ultrastructure was similar in monolayers and spheroids, and only from the fifth day in Culture, did hepatocytes in the centre of floating aggregates become necrotic. From day 1 to day 5, hepatocytes in Primary Culture displayed only minor cytological alterations. Excellent cytoplasmic compartmentation, restoration of hepatocytic glycogen stores, high secretion rates of very low density lipoproteins by dictyosomes, establishment of cell-to-cell contacts, restitution of cellular polarity and the epithelial character of the cells, as well as formation of bile canaliculi documented recovery of the hepatocytes in Primary Culture. From day 5 in Culture, an increasing number of cells detached from the substratum, and cell senescence was indicated by a marked increase in ultrastructural heterogeneity, with progressive vesiculation and fractionation of the RER, transformation of RER stacks into huge membrane whorls, aggregation and proliferation of peroxisomes and SER, lack of dictyosomal VLDL production, drastic accumulation of lysosomes, myelinated bodies and autophagic vacuoles, as well as successive exhaustion of cellular glycogen deposits. Whereas with conventional methods for assessing cell viability, most hepatocytes appeared intact for up to ten days in Culture, cytological investigations revealed severe deterioration of cellular integrity from day 7. However, for incubation periods of up to five days, isolated rainbow trout hepatocytes can be recommended as an excellent model for physiological and toxicological studies.
Jun Han - One of the best experts on this subject based on the ideXlab platform.
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Primary Culture of Porcine Pancreatic Acinar Cells
Pancreas, 2002Co-Authors: Chengwei Tang, Xudong Zhao, Jun HanAbstract:Objective To develop a method for the Primary Culture of porcine pancreatic acinar cells. Interventions Dispersed pancreatic acinar cells available utilizing RPMI-1640 medium containing collagenase III. After purification, the isolated acinar cells were Cultured in RPMI1640 medium with the addition of 2.5% fetal bovine serum. Main outcome measures The morphological characteristics of acinar cells were described. 3 H-thymidine incorporation of acinar cells and the activity of amylase or lipase were determined during the Culture process. Results There were no remarkable morphological changes in the pancreatic acinar cells during the 20 days ’ Culture. The acini showed a tendency to gather but did not attach to the walls of the Culture disks. A good 3 Hthymidine incorporation of acinar cells in the Primary Culture was maintained. The secretion of amylase or lipase from the acini decreased with the length of time of the Culture. Discussion The Primary Culture of acinar cells from a porcine pancreas which was carried out in this study maintained the normal morphology of the acinar cells and the ir ability to grow but not the ir secretion of amylase or lipase. The method would benefit by the further experiments on acini of porcine pancreas.
T. Braunbeck - One of the best experts on this subject based on the ideXlab platform.
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Senescence of hepatocytes isolated from rainbow trout (Oncorhynchus mykiss) in Primary Culture
Protoplasma, 1992Co-Authors: T. Braunbeck, V. StorchAbstract:In order to evaluate the suitability of isolated fish hepatocytes for toxicological studies, hepatocytes were isolated from rainbow trout ( Oncorhynchus mykiss ) by liver collagenase perfusion. Isolated hepatocytes were investigated for seven days in Primary Culture on uncoated petri dishes using light and electron microscopy. Viability of isolated hepatocytes, as estimated from trypan blue exclusion, declined from >90% at the beginning of the incubation to ⩽80% after eight days of Primary Culture. Survival of hepatocytes was best at an incubation temperature of 14°C, and addition of fetal calf serum failed to improve cell performance. Freshly isolated hepatocytes appeared as solitary spherical cells with numerous microvilli at the outer surface. Except for a 30% reduction in cell size due to stress-induced glycogen reduction, the ultrastructure of freshly isolated hepatocytes closely resembled that of rainbow trout hepatocytes in vivo, which is characterized by distinct cytoplasmic segregation into a perinuclear portion containing rough endoplasmic reticulum (RER) arranged in extensive stacks, mitochondria, peroxisomes and the peribiliary complex (dictyosomes, lysosomes), and large peripheral glycogen fields occasionally interspersed with small lipid inclusions. Within one day of Culture, about 60–80% of the isolated hepatocytes sedimented to form a monolayer attached to the Culture dishes, whereas up to 20% remained in suspension forming hepatocyte aggregates. Cell adhesion was weak, and during prolonged Culture increasing amounts of cell detached, whereas the floating cell accumulations grew to aggregates of more than 100 cells. Cell viability and ultrastructure was similar in monolayers and spheroids, and only from the fifth day in Culture, did hepatocytes in the centre of floating aggregates become necrotic. From day 1 to day 5, hepatocytes in Primary Culture displayed only minor cytological alterations. Excellent cytoplasmic compartmentation, restoration of hepatocytic glycogen stores, high secretion rates of very low density lipoproteins by dictyosomes, establishment of cell-to-cell contacts, restitution of cellular polarity and the epithelial character of the cells, as well as formation of bile canaliculi documented recovery of the hepatocytes in Primary Culture. From day 5 in Culture, an increasing number of cells detached from the substratum, and cell senescence was indicated by a marked increase in ultrastructural heterogeneity, with progressive vesiculation and fractionation of the RER, transformation of RER stacks into huge membrane whorls, aggregation and proliferation of peroxisomes and SER, lack of dictyosomal VLDL production, drastic accumulation of lysosomes, myelinated bodies and autophagic vacuoles, as well as successive exhaustion of cellular glycogen deposits. Whereas with conventional methods for assessing cell viability, most hepatocytes appeared intact for up to ten days in Culture, cytological investigations revealed severe deterioration of cellular integrity from day 7. However, for incubation periods of up to five days, isolated rainbow trout hepatocytes can be recommended as an excellent model for physiological and toxicological studies.
