Mesenchymal Stromal Cell on Liver Decellularised Extracellular Matrix for Tissue Engineering.

BACKGROUND: In end-stage chronic liver disease, transplantation represents the only curative option. However, the shortage of donors results in the death of many patients. To overcome this gap, it is mandatory to develop new therapeutic options. In the present study, we decellularised pig livers and reseeded them with allogeneic porcine mesenchymal stromal cells (pMSCs) to understand whether extracellular matrix (ECM) can influence and/or promote differentiation into hepatocyte-like cells (HLCs). METHODS: After decellularisation with SDS, the integrity of ECM-scaffolds was examined by histological staining, immunofluorescence and scanning electron microscope. DNA quantification was used to assess decellularisation. pMSCs were plated on scaffolds by static seeding and maintained in in vitro culture for 21 days. At 3, 7, 14 and 21 days, seeded ECM scaffolds were evaluated for cellular adhesion and growth. Moreover, the expression of specific hepatic genes was performed by RT-PCR. RESULTS: The applied decellularisation/recellularisation protocol was effective. The number of seeded pMSCs increased over the culture time points. Gene expression analysis of seeded pMSCs displayed a weak induction due to ECM towards HLCs. CONCLUSIONS: These results suggest that ECM may address pMSCs to differentiate in hepatocyte-like cells. However, only contact with liver-ECM is not enough to induce complete differentiation.

In-Lab Manufacturing of Decellularized Rat Renal Scaffold for Kidney Bioengineering.

Whole-organ decellularization is recently gaining interest in the transplantation field as strategy to obtain acellular scaffold only composed by extracellular matrix. These structures, that still remain organ-specific in terms of biological cues and tridimensional morphology could be then recellularized with patient's autologous cells. The final result should be a nwe transplantable autologous organ that should by-pass, at the same time, the problem of organ shortage and secondly the consequences related to the immunosuppression need. Herein we describe the protocol to manufacture a whole-organ transplantable rat kidney scaffold by a dual-detergent (Triton X-100 and SDS) arterial peristaltic perfusion. Final results show whole-renal acellular scaffold with contextual preservation of tridimensional architecture and biological properties deriving from the extracellular matrix composition.