Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic Wounds

Diabetic foot ulcers (DFUs) are associated with a high risk of amputations and a 50% 5-year survival rate due at least in part to the limited angiogenic and wound healing capacity of patients with diabetes. Cell therapy via intramuscular injection of peripheral blood mononuclear cells showed encoura...

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Main Authors: Andrea Rampin, Carlo Maria Ferdinando Caravaggi, Luigi Troisi, Gaia Spinetti
Format: Article
Language:English
Published: SAGE Publishing 2025-03-01
Series:Cell Transplantation
Online Access:https://doi.org/10.1177/09636897241309698
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author Andrea Rampin
Carlo Maria Ferdinando Caravaggi
Luigi Troisi
Gaia Spinetti
author_facet Andrea Rampin
Carlo Maria Ferdinando Caravaggi
Luigi Troisi
Gaia Spinetti
author_sort Andrea Rampin
collection DOAJ
description Diabetic foot ulcers (DFUs) are associated with a high risk of amputations and a 50% 5-year survival rate due at least in part to the limited angiogenic and wound healing capacity of patients with diabetes. Cell therapy via intramuscular injection of peripheral blood mononuclear cells showed encouraging but limited results. Such limitations may arise from the limited ability of therapeutic cells to adhere to the target tissue. The development of a methodology able to support the targeted delivery of viable angiogenic cells would improve cell therapy outcomes in DFU. Here, we optimized a protocol for the production of autologous extracellular matrix (ECM)-rich pericyte-based cell sheets for cell delivery. Pericytes were isolated from skeletal muscle biopsies of DFU patients and non-diabetic controls and characterized by flow cytometry and immunofluorescence. Human umbilical vein endothelial cells used for the optimization of collagen IV deposition showed a positive correlation with seeding density and a negative one with sub-culture passaging ( P < 0.05). Macromolecular crowding significantly increased collagen IV deposition both in human umbilical vein endothelial cells and in patient-derived pericytes ( P < 0.01) without affecting proliferation ( P > 0.05). Finally, DFU patient-derived pericytes effectively deposited ECM supporting their use for autologous cell sheet production.
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spelling doaj-art-192e1d2fa05a4ec1a21dca106159816b2025-08-20T02:57:35ZengSAGE PublishingCell Transplantation1555-38922025-03-013410.1177/09636897241309698Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic WoundsAndrea Rampin0Carlo Maria Ferdinando Caravaggi1Luigi Troisi2Gaia Spinetti3Cardiovascular Research Unit, IRCCS MultiMedica, Milan, ItalyDiabetic Foot Department, IRCCS MultiMedica, Milan, ItalyReconstructive Microsurgery Service – Department of Hand Surgery & Rehabilitation-IRCCS MultiMedica, Milan, ItalyCardiovascular Research Unit, IRCCS MultiMedica, Milan, ItalyDiabetic foot ulcers (DFUs) are associated with a high risk of amputations and a 50% 5-year survival rate due at least in part to the limited angiogenic and wound healing capacity of patients with diabetes. Cell therapy via intramuscular injection of peripheral blood mononuclear cells showed encouraging but limited results. Such limitations may arise from the limited ability of therapeutic cells to adhere to the target tissue. The development of a methodology able to support the targeted delivery of viable angiogenic cells would improve cell therapy outcomes in DFU. Here, we optimized a protocol for the production of autologous extracellular matrix (ECM)-rich pericyte-based cell sheets for cell delivery. Pericytes were isolated from skeletal muscle biopsies of DFU patients and non-diabetic controls and characterized by flow cytometry and immunofluorescence. Human umbilical vein endothelial cells used for the optimization of collagen IV deposition showed a positive correlation with seeding density and a negative one with sub-culture passaging ( P < 0.05). Macromolecular crowding significantly increased collagen IV deposition both in human umbilical vein endothelial cells and in patient-derived pericytes ( P < 0.01) without affecting proliferation ( P > 0.05). Finally, DFU patient-derived pericytes effectively deposited ECM supporting their use for autologous cell sheet production.https://doi.org/10.1177/09636897241309698
spellingShingle Andrea Rampin
Carlo Maria Ferdinando Caravaggi
Luigi Troisi
Gaia Spinetti
Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic Wounds
Cell Transplantation
title Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic Wounds
title_full Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic Wounds
title_fullStr Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic Wounds
title_full_unstemmed Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic Wounds
title_short Macromolecular Crowding Supports the Generation of Basal Membrane-Rich Pericyte-Based Cell Sheets Useful for Cell Therapy of Diabetic Wounds
title_sort macromolecular crowding supports the generation of basal membrane rich pericyte based cell sheets useful for cell therapy of diabetic wounds
url https://doi.org/10.1177/09636897241309698
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