Heparin Anti-liver Fibrosis And Clinical Studies

Objective This section was aimed to establish the method of separation ofhepatic stellate cells (HSCs) and Kupffer cells (KCs) simultaneously.Method Non-parenchymal cells were fractionated by sequential collagenase and pronase digestion of liver. 18% (W/V ) Nycodenz was used to separate HSCs and KCs. HSCs were present at the boundary,while KCs were present below the boundary. Viability of HSCs and KCs were measured by the Trypan blue exclusion test. HSCs were established by the visualization of desmin with anti-desmin antibody by immunocytochemistry staining.KCs were established by the visualization of lysozyme by immunocytochemistry staining.Results Purity of these cultures was about 95%.This method for cell isolation resulted in a yield of 3.0±0.5×10~7 HSCs and 4.0 ± 0.5×10~6 KCs per liver.Conclusion HSCs and KCs can be separated by one-step density gradient centrifuge with nycodenz.