In Vitro Conditioning of Adipose-Derived Mesenchymal Stem Cells by the Endothelial Microenvironment: Modeling Cell Responsiveness towards Non-Genetic Correction of Haemophilia A

Abstract

In recent decades, the use of adult multipotent stem cells has paved the way for the identification of new therapeutic approaches for the treatment of monogenic diseases such as Haemophilia A. Being already studied for regenerative purposes, adipose-derived mesenchymal stem cells (Ad-MSCs) are still poorly considered for Haemophilia A cell therapy and their capacity to produce coagulation factor VIII (FVIII) after proper stimulation and without resorting to gene transfection. In this work, Ad-MSCs were in vitro conditioned towards the endothelial lineage, considered to be responsible for coagulation factor production. The cells were cultured in an inductive medium enriched with endothelial growth factors for up to 21 days. In addition to significantly responding to the chemotactic endothelial stimuli, the cell populations started to form capillary-like structures and up-regulated the expression of specific endothelial markers (CD34, PDGFRα, VEGFR2, VE-cadherin, CD31, and vWF). A dot blot protein study detected the presence of FVIII in culture media collected from both unstimulated and stimulated Ad-MSCs. Remarkably, the activated partial thromboplastin time test demonstrated that the clot formation was accelerated, and FVIII activity was enhanced when FVIII deficient plasma was mixed with culture media from the untreated/stimulated Ad-MSCs. Overall, the collected evidence supported a possible Ad-MSC contribution to HA correction via specific stimulation by the endothelial microenvironment and without any need for gene transfection.

Identification of the Efficient Enhancer Elements in FVIII-Padua for Gene Therapy Study of Hemophilia A

“…They are considered ideal therapeutic cells for many diseases owing to their low immunogenicity, strong proliferative ability, wide and easy-to-obtain tissue sources, multi-directional differentiation potential, and homing and paracrine effect [34][35][36][37][38]. Under physiological conditions, MSCs synthesize and secrete FVIII [39] and differentiate into ECs [40]. Co-transplantation of MSCs and ECs resulting in enhanced cell engraftment and FVIII expression in mice [41,42].…”

Editorial: Therapeutic potential of mesenchymal stem cells in organ and tissue regeneration

“…Additionally, MSCs lack the expression of histocompatibility complexes and immune-stimulating molecules, and as they are not detected by host immune surveillance, their use reduces graft rejection after transplantation. For these reasons, MSCs have turned out to be competent candidates in tissue engineering ( Han et al, 2019 ; Barbon et al, 2022 ).…”

In Vitro Models of Tissue and Organ Regeneration

“…Two original works use in vitro models of mesenchymal stromal/stem cells (MSCs) to investigate regenerative purposes. Barbon and coworkers use an in vitro conditioning regimen of MSCs towards the endothelial lineage to stimulate coagulation factor VIII production [5]. The background of this work was the development of a cell therapy for the treatment of Haemophilia A and, therefore, for future pre-clinical investigation using preconditioned MSCs.…”