An approach to preparing decellularized whole liver organ scaffold in rat
Issue title: Selected Papers from the 5th China–France International Symposium “Stem Cells: From Bench to Bedside”, 12–14 December 2013, Kunming, China
Affiliations: BRC, First Hospital of Kun Ming, Kun Ming, China | Lorraine University and CNRS UNR 7561, Medical College, Vandœuvre-lès-Nancy, France | CHRU Nancy, Unité Therapie Cellulaire et Tissulaire, Vandœuvre-lès-Nancy, France
Note: [] Address for correspondence: Lei Zhang, Biomedical Research Center, Affiliated Calmette Hospital of Kun Ming Medical University, Yun Nan, China. E-mail: zlei01@hotmail.com
Abstract: OBJECTIVES: In present study, we plan to produce a decellularization protocol from rat liver to generate a three-dimensional whole organ scaffold. METHODS: A combination of 1% SDS and 1% tritonX-100 were used orderly to decellularize rat livers. After about 6 h of interactive antegrade/retrograde perfusion, a decellularized whole translucent liver scaffold with integrated blood vessel networks was generated. The decellularized livers are charactered by light microscopy, scanning electron microscopy, and biochemical analysis (DNA quantification) for preservation of the three-dimension of extracellular matrix architecture. RESULTS: The decellularization protocol was verified by observation of the whole translucent liver organ with intact vascular trees under macroscopy, in conjunction with the hematoxylin–eosin staining that showed no cells or nuclear material remained. Additionally, the Masson's stain indicted that the extracellular proteins were well kept and scanning electron microscopy (SEM) revealed a preserved decellularized matrix architecture. Compared to normal livers, DNA in the decellularized livers was quantified less than 10% at the same mass. CONCLUSIONS: The current method of decellularization protocol was feasible, simple and quick, and was verified by an absence of residual cells. The decellularized extracellular matrix had preserved integrate vascular network and a three-dimensional architecture.
Keywords: Decellularization, perfusion, whole liver organ scaffold
