Using a decellularized splenic matrix as a 3D scaffold for hepatocyte cultivation in vitro : a preliminary trial

Abstract

Using a decellularized liver matrix (DLM) to reengineer liver tissue is a promising therapy for end-stage liver disease. However, the limited supply of donor organs still hampers its potential clinical application, while a xenogenic decellularized matrix may bring a risk of zoonosis and immunological rejection. Therefore, an appropriate alternative scaffold is needed. In this research, we established a decellularized splenic matrix (DSM) in a rodent model, which preserved the 3D ultrastructure, the components of the extracellular matrix (ECM) and the native vascular network. The DSM and DLM had similar components of ECM, and similar mechanical properties. Hepatocytes were seeded to the DSM and DLM for dynamic culturing up to 6 d, and distributed both in decellularized sinusoidal spaces and around the vessels. The TUNEL-positive cell percentage in a dynamic culturing decellularized splenic matrix (dDSM) was 10.7% ± 3.6% at 3d and 25.8% ± 5.6% at 5d, although 14.2% ± 4.5% and 24.8% ± 2.9%, respectively, in a dynamic culturing decellularized liver matrix (dDLM) at the same time point (p > 0.05). Primary hepatocytes in the dDSM and dDLM expressed albumin, G6pc and Ugt1a1. The gene expression of Cyp2b1, Cyp1a2 and HNF1α in the gene transcription level revealed hepatocytes had lower gene expression levels in the dDSM compared with the dDLM at 3d, but better than those in a sandwich culture. The cumulative albumin production at 6 d of culture was 80.7 ± 9.6 μg per million cells in the dDSM and 89.6 ± 4.6 μg per million cells in the dDLM (p > 0.05). In summary, the DSM is a promising 3D scaffold for hepatocyte cultivation in vitro.

Decellularized spleen matrix for reengineering functional hepatic-like tissue based on bone marrow mesenchymal stem cells

“…Remarkably, an abundant blood sinus structure was observed in DSM, which was favorable to satisfy the large amount of cell metabolism and oxygen consumption. It has been concluded in our published reports 15,16 that the hepatocyte vitality when cultured in DSM as well as the synthesis and secretion function were comparable with DLM-based HLT.…”

“…Herein, in order to expand the scaffold donor pool, we have established the decellularized spleen matrix (DSM), which had homogeneous ECM with DLM, and presented comparable condition for hepatocyte cultivation. 15,16 DSM scaffolds may be more promising for clinical application due to their extensive sources such as traumatic rupture, hypersplenism caused by various reasons, idiopathic thrombocytopenic purpura, floating spleen, and donation after cardiac/ brain death.…”

Strategies based on organ decellularization and recellularization

“…However, an organ might be reconstructed by cultivating organ‐specific cells in an ECM derived from another organ (e.g. spleen and hepatocytes or hepatocyte‐like cells) . Although this concept promises to be very practical, it only appears to be suitable for organs possessing an endocrine function (e.g.…”

The effect of riboflavin/UVA cross‐linking on anti‐degeneration and promoting angiogenic capability of decellularized liver matrix

“…We established DLM scaffolds by in a rodent model. Our previous work described the characterization of decellularized liver and spleen matrix . Briefly, by decellularization, the gross morphology of liver was well‐preserved with visible vascular structure.…”

“…Our previous work described the characterization of decellularized liver and spleen matrix. 6,14 Briefly, by decellularization, the gross morphology of liver was well-preserved with visible vascular structure. HE, DAPI nuclear staining and remnant DNA quantitative analysis of the DLM confirmed the absence of cellular components and nuclear material.…”