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The Isolation of Kupffer Cells in BALB/c Mouse

Funan Qiu

Open publisher page 1 citations

Abstract

Objective To explore a better method for isolation of mouse Kupffer cells(KC).Methods KC was isolated from the liver of mouse by in situ perfusion with enzyme,enzymatic digestion ex vivo,discontinuous density gradient centrifugation and selective adherence,we compared pronase E,collagenase Ⅳ,pronase E and collagenase Ⅳ mix for isolation of KC in yield and purity.Results The yield of KC with three different methods was(6.32±0.5)×106 g-1,(3.66±0.4)×106 g-1,(10.3±0.7)×106 g-1,respectively.The purity was(93.2±1.7)%,(90.7±1.5)%,(94.5±1.9)%,respectively.Conclusion Combining pronase E with collagenase Ⅳ in situ perfusion and enzymatic digestion ex vivo may be a better method for isolation of KC.

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Objective To explore a better method for isolation of mouse Kupffer cells(KC).Methods KC was isolated from the liver of mouse by in situ perfusion with enzyme,enzymatic digestion ex vivo,discontinuous density gradient centrifugation and selective adherence,we compared pronase E,collagenase Ⅳ,pronase E and collagenase Ⅳ mix for isolation of KC in yield and purity.Results The yield of KC with three different methods was(6.32±0.5)×106 g-1,(3.66±0.4)×106 g-1,(10.3±0.7)×106 g-1,respectively.The purity was(93.2±1.7)%,(90.7±1.5)%,(94.5±1.9)%,respectively.Conclusion Combining pronase E with collagenase Ⅳ in situ perfusion and enzymatic digestion ex vivo may be a better method for isolation of KC.

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Available abstract

Objective To explore a better method for isolation of mouse Kupffer cells(KC).Methods KC was isolated from the liver of mouse by in situ perfusion with enzyme,enzymatic digestion ex vivo,discontinuous density gradient centrifugation and selective adherence,we compared pronase E,collagenase Ⅳ,pronase E and collagenase Ⅳ mix for isolation of KC in yield and purity.Results The yield of KC with three different methods was(6.32±0.5)×106 g-1,(3.66±0.4)×106 g-1,(10.3±0.7)×106 g-1,respectively.The purity was(93.2±1.7)%,(90.7±1.5)%,(94.5±1.9)%,respectively.Conclusion Combining pronase E with collagenase Ⅳ in situ perfusion and enzymatic digestion ex vivo may be a better method for isolation of KC.

Key concepts: Pronase, Collagenase, Ex vivo, Digestion (alchemy), In vivo, Centrifugation, Chemistry, Molecular biology

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