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The Investigation of Isolation, Culture, Characterization and Activation of Pancreatic Stellate Cells
Author: ZhouLongAn
Tutor: QianZhuYin
School: Nanjing Medical University
Course: General Surgery
Keywords: Pancreatic stellate cells Abdominal aortic Perfusion Culture Appraisal Smad2 / 3
CLC: R329
Type: Master's thesis
Year: 2008
Downloads: 79
Quote: 0
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Abstract
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Objective: abdominal aortic infusion collagenase digestion and separation of pancreatic stellate cells ( pancreatic stellate cell PSC) to establish simple PSC separation method , and the culture and identification of PSC ; explore the the PSC early activation mechanism . Methods : Rat abdominal aortic perfusion intubation by collagenase digestion Nycodenz density gradient centrifugation , isolated PSC ; by cell morphology was observed lipid droplets within the cytoplasm and immunocytochemistry method detection desmin ( desmin, ) , glial quality acid fibrin ( the glial fibrillaryacidic protein , GFAP ) , alpha smooth muscle actin hormone (α-smooth muscle actin, α-SMA) expression identification PSC ; Determination of PSC Smad2 / 3 expression by RT- PCR method and WestBlot of methods . Results: abdominal aortic perfusion separation PSC, yield , vigor and purity were : 15.3 ± 4.6 × 10 ~ 3 / g body weight , 95.0 ± 3.5 % , > 80% ; most cells after 24h incubation adherent star-shaped or polygonal 48 hours after the expression of desmin, of GFAP expression of α-SMA and after 96h ; Smad2 / 3 the PSC early activation significantly high expression reached its peak after 48H . Conclusion: easier perfusion through the abdominal aorta isolated PSC , the higher its yield , vigor and purity , was identified to meet the requirements of the in vitro experiments ; Smad2 / 3 expression is the central link in the the PSC early activation process . Smad2 / 3 high expression of Smad7 can not inhibit its activation resulting in the loss of balance between the two is the main reason for PSC activation .
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CLC: > Medicine, health > Basic Medical > Human morphology > Human histology
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