Obtaining liver tridimensional scaffold through the decellularization of rabbit whole liver in 24 hours

Abstract

In the present work, we development a new protocol for liver decellularization in which the hole decellularization was reached over 24 h.

Introduction: the availability of transplantable livers is not sufficient to fulfill the current demand for grafts, with the search for therapeutic alternatives having generated different lines of research, one of which is the use of decellularized three-dimensional biological matrices and subsequent cell seeding to obtain a functional organ.

Objective: to produce a decellularization protocol from rabbit liver to generate a three-dimensional matrixin which the time period involved didn't pass 24 h.

Methods: The decellularization is obtained through the use of water and SDS (0,1-0,3 %), after freezing at -80 degrees, is the best alternative of different physical and/or chemical mechanisms to break down organ cells and leave only the extracellular matriz. After 24 h of retrograde perfusion, a decellularized translucent matrix was generated. To evaluate if the decellularization protocol was successful, with the extracellular matrix being preserved, we carried out histological (light microscopy) and biochemical (DNA quantification) studies.

Results: the decellularization process was verified by macroscopic observation of the organ using microscopic observation corroborated the macroscopic results, with the hematoxylin-eosin and Masson staining showing no cells or nuclear material. In addition, the DNA quantification was less than 10% in the decellularized liver compared to control. Finally,the time taken to develop the decellularization protocol was less than 24 hours.