Determining the ideal decellularization protocol applied for preparing for liver biological scaffold / 重庆医学

ABSTRACT

Objective To develop an appropriate and simple decellularization protocol for preparing decellularized liver bio-logical scaffold(DLBS) .Methods Through immune-ofluorescence analysis ,scanning electron microscope ,quantification of Glyco-samin-oglycans(GAGs) and DNA ,we evaluated the effectiveness of low concentrations of SDS(0 .25% ,0 .50% ) and 1% Triton X-100 in the preservation of extracellular matrix of rat livers .Then ,we assessed Cytotoxicity of DLBS treated with three methods by MTT assay .Results Residual DNA after SDS(0 .25% ,0 .50% ) treatments were below 50 ng/mg dry weight ,which were less than Triton X-100 treatments .The content of GAGs in the 0 .25% SDS and 1% Triton X-100-treated scaffolds were higher than other detergent-treated scaffolds .The result of MTT assay showed that the liver scaffolds treated with three methods were not cytotoxic to proliferation of C3A cells .Conclusion The protocol containing 0 .25% SDS and a perfusion rate of 5 mL/min could be a rapid and effective decellularization protocol .DLBS constructed using this protocol could be an ideal material for preparing a transplant-able organ .