Multi-stage bioengineering of a layered oesophagus with in vitro expanded muscle and epithelial adult progenitors

Abstract

A tissue engineered oesophagus could overcome limitations associated with oesophageal substitution. Combining decellularized scaffolds with patient-derived cells shows promise for regeneration of tissue defects. In this proof-of-principle study, a two-stage approach for generation of a bio-artificial oesophageal graft addresses some major challenges in organ engineering, namely: (i) development of multi-strata tubular structures, (ii) appropriate re-population/maturation of constructs before transplantation, (iii) cryopreservation of bio-engineered organs and (iv) in vivo pre-vascularization. The graft comprises decellularized rat oesophagus homogeneously re-populated with mesoangioblasts and fibroblasts for the muscle layer. The oesophageal muscle reaches organised maturation after dynamic culture in a bioreactor and functional integration with neural crest stem cells. Grafts are pre-vascularised in vivo in the omentum prior to mucosa reconstitution with expanded epithelial progenitors. Overall, our optimised two-stage approach produces a fully re-populated, structurally organized and pre-vascularized oesophageal substitute, which could become an alternative to current oesophageal substitutes.

Patient-Derived Scaffolds of Colorectal Cancer Metastases as an Organotypic 3D Model of the Liver Metastatic Microenvironment

“…The cell migration capability was also analyzed in patient-derived scaffolds by measuring bioluminescence at the far end of scaffolds and after a short culture period to minimize the contribution from cell proliferation [23,24]. We identified a significantly enhanced migration and invasion capability in CRC cells grown in CRLM scaffolds, compared with the healthy counterpart.…”

Engineering transplantable jejunal mucosal grafts using primary patient-derived organoids from children with intestinal failure

“…The DET decellularization protocol represents a more physiological approach for the fabrication of non-immunogenic biological scaffolds as compared to synthetic polymers. The ability to preserve the biological and structural features of the decellularised scaffolds after cryopreservation further suggests the potential of these scaffolds to become “off-shelf” clinical products 13 . Previous studies have demonstrated the conservation of the biological ECM molecules (e.g.…”

“…Autologous tissue engineering of functional intestinal grafts, through combination of biomaterials and patient-derived cells, presents an innovative alternative approach to treat IF patients 6 . Multiple studies have previously reported the pre-clinical developments of tissue engineering methodologies in other organs such as oesophageal, skeletal muscle, liver and lung reconstruction 7-13 . However, while engineering of simpler tissues such as skin and cornea are established in clinical practice 14,15 examples of successful clinical applications of more complex organs have only been demonstrated in few case reports of airway and bladder reconstruction 16,17 .…”

Isolation and characterisation of mouse intestinal mesoangioblasts

“…When derived from adult tissue, MABs have been shown to generate both vascular and other mesodermal derivatives [18] including contributing to the regeneration of the neuromuscular wall of a bioengineered oesophagus [10]. Given this regenerative potential it is possible that MABs may provide an ideal cell source for regeneration of intestinal smooth muscle cells.…”