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- W2393219483 abstract "Objective: To establish a simple and reliable technique to isolate and culture the rat hepatocytes and kupffer cells simultaneously.Methods: To obtain liver cell suspension,the liver in vivo was perfused at first,then the liver with collagenase in vitro were perfused and digested.The filtrated cell suspension was separated into liver parenchymal and non-parenchymal cells using differential centrifugation.Then hepatocytes were harvested by repeatedly low speed centrifugation,kupffer cells were purified by density gradient centrifugation with percoll and further enriched by selective attachment.The viability of these cells was determined by Typan blue exclusion staining.Inverted phase contrast microscope,HE staining,periodic acid-schiff(PAS) staining and immunohistochemical staining of albumin were used to examine the morphological characteristics and function of the hepatocytes.Kupffer cells were identificated by light microscope,fluorescent microscope and immunofluorescence staining of CD68.Results: Hepatocytes and kupffer cells were isolated synchronously and cultured successfully in vitro.The yield of hepatocytes was about 1.37±0.53×108 per rat and 3.45±0.41×106 kupffer cells were harvested per gram liver.The viability and purity of these cells were 90%.Hepatocytes presented its typical cell morphology after 24 hours of cultivation and still remained functions of glycogen and albumin synthesis after day 7.The isolated kupffer cells showed polymorphism with typical polygon-like and star-like shapes and exhibited positive result of CD68.Conclusion: Hepatocytes and kupffer cells with high viability and function can be separated by the modified perfusion technique." @default.
- W2393219483 created "2016-06-24" @default.
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- W2393219483 date "2013-01-01" @default.
- W2393219483 modified "2023-09-24" @default.
- W2393219483 title "Establishment of a Technique to Isolate and Culture of Rat Hepatocytes and Kupffer Cells Simultaneously" @default.
- W2393219483 hasPublicationYear "2013" @default.
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