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Publications

Journal Article Research Support, Non-U.S. Gov't Epublish 3 citations

PubMed: 37880340 DOI: 10.1038/s41598-023-45610-5

Development and qualification of clinical grade decellularized and cryopreserved human esophagi.

Sci Rep 13(1):18283

Authors: William Godefroy, Lionel Faivre, Caroline Sansac, Briac Thierry, Jean-Marc Allain, Patrick Bruneval, Rémy Agniel, Sabrina Kellouche, Olivier Monasson, Elisa Peroni, Mohamed Jarraya, Niclas Setterblad, Massymissa Braik, Benjamin Even, Sophie Cheverry, Thomas Domet, Patricia Albanese, Jérôme Larghero, Pierre Cattan, Lousineh Arakelian

Affiliations

  • Service de Chirurgie Viscérale, Cancérologique et Endocrinienne, Hôpital Saint-Louis - Université Paris Cité, Paris, France.
  • Unité de Thérapie Cellulaire, Hôpital Saint-Louis, AP-HP, Paris, France.
  • CIC de Biothérapies CBT 501, Paris, France.
  • Human Immunology, Pathophysiology, Immunotherapy / HIPI / INSERM UMR976, Laboratoire de Biotechnologies de Cellules Souches, Université Paris Cité, 75010, Paris, France.
  • Unité de Thérapie Cellulaire, Hôpital Saint-Louis, AP-HP, Paris, France.
  • CIC de Biothérapies CBT 501, Paris, France.
  • Human Immunology, Pathophysiology, Immunotherapy / HIPI / INSERM UMR976, Laboratoire de Biotechnologies de Cellules Souches, Université Paris Cité, 75010, Paris, France.
  • Banque de Tissus Humains, Hôpital St-Louis, AP-HP, Paris, France.
  • Service d'ORL Pédiatrique, AP-HP, Hôpital Universitaire Necker, 75015, Paris, France.
  • LMS, CNRS, Ecole Polytechnique, Institut Polytechnique de Paris, Palaiseau, France.
  • Inria, Paris, France.
  • Service d'Anatomie Pathologie, Hôpital Européen Georges Pompidou, AP-HP, Paris, France.
  • Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des Matériaux, I-MAT (FD4122), CY Cergy Paris Université, Cergy-Pontoise, France.
  • CNRS, BioCIS, CY Cergy Paris Université, 95000, Cergy Pontoise, France.
  • CNRS, BioCIS, Université Paris-Saclay, 92290, Châtenay-Malabry, France.
  • UMS Saint-Louis US53 / UAR2030, Institut de Recherche Saint-Louis Plateforme Technologique Centre, Université Paris Cité - Inserm - CNRS, Paris, France.
  • Laboratoire Gly-CRRET, Université Paris Est Créteil, Université Paris Est, EA 4397 ERL CNRS 9215, Créteil, France.
  • Centre MEARY de Thérapie Cellulaire Et Génique, AP-HP, Hôpital Saint-Louis, 75010, Paris, France.
  • Service de Chirurgie Viscérale, Cancérologique et Endocrinienne, Hôpital Saint-Louis - Université Paris Cité, Paris, France.
  • Unité de Thérapie Cellulaire, Hôpital Saint-Louis, AP-HP, Paris, France.
  • CIC de Biothérapies CBT 501, Paris, France.
  • Human Immunology, Pathophysiology, Immunotherapy / HIPI / INSERM UMR976, Laboratoire de Biotechnologies de Cellules Souches, Université Paris Cité, 75010, Paris, France.

Abstract

Tissue engineering is a promising alternative to current full thickness circumferential esophageal replacement methods. The aim of our study was to develop a clinical grade Decellularized Human Esophagus (DHE) for future clinical applications. After decontamination, human esophagi from deceased donors were placed in a bioreactor and decellularized with sodium dodecyl sulfate (SDS) and ethylendiaminetetraacetic acid (EDTA) for 3 days. The esophagi were then rinsed in sterile water and SDS was eliminated by filtration on an activated charcoal cartridge for 3 days. DNA was removed by a 3-hour incubation with DNase. A cryopreservation protocol was evaluated at the end of the process to create a DHE cryobank. The decellularization was efficient as no cells and nuclei were observed in the DHE. Sterility of the esophagi was obtained at the end of the process. The general structure of the DHE was preserved according to immunohistochemical and scanning electron microscopy images. SDS was efficiently removed, confirmed by a colorimetric dosage, lack of cytotoxicity on Balb/3T3 cells and mesenchymal stromal cell long term culture. Furthermore, DHE did not induce lymphocyte proliferation in-vitro. The cryopreservation protocol was safe and did not affect the tissue, preserving the biomechanical properties of the DHE. Our decellularization protocol allowed to develop the first clinical grade human decellularized and cryopreserved esophagus.

MeSH

Cryopreservation, Balb 3T3 Cells