Highly Angiogenic, Nonthrombogenic Bone Marrow Mononuclear Cell–Derived Spheroids in Intraportal Islet Transplantation

Abstract

Highly angiogenic bone marrow mononuclear cell-derived spheroids (BM-spheroids), formed by selective proliferation of the CD31CD14CD34 monocyte subset via three-dimensional (3D) culture, have had robust angiogenetic capacity in rodent syngeneic renal subcapsular islet transplantation. We wondered whether the efficacy of BM-spheroids could be demonstrated in clinically relevant intraportal islet transplantation models without increasing the risk of portal thrombosis. The thrombogenic potential of intraportally infused BM-spheroids was compared with that of mesenchymal stem cells (MSCs) and MSC-derived spheroids (MSC-spheroids). The angiogenic efficacy and persistence in portal sinusoids of BM-spheroids were examined in rodent syngeneic and primate allogeneic intraportal islet transplantation models. In contrast to MSCs and MSC-spheroids, intraportal infusion of BM-spheroids did not evoke portal thrombosis. BM-spheroids had robust angiogenetic capacity in both the rodent and primate intraportal islet transplantation models and improved posttransplant glycemic outcomes. MRI and intravital microscopy findings revealed the persistence of intraportally infused BM-spheroids in portal sinusoids. Intraportal cotransplantation of allogeneic islets with autologous BM-spheroids in nonhuman primates further confirmed the clinical feasibility of this approach. In conclusion, cotransplantation of BM-spheroids enhances intraportal islet transplantation outcome without portal thrombosis in mice and nonhuman primates. Generating BM-spheroids by 3D culture prevented the rapid migration and disappearance of intraportally infused therapeutic cells.

Intraportally delivered stem cell spheroids localize in the liver and protect hepatocytes against GalN/LPS-induced fulminant hepatic toxicity

“…In a recent study, thrombosis and infarction were more prominent after intraportal delivery of 2D-ADSC compared to the 3D-ADSC-delivered group [42], which may be due to higher expressions of thrombogenic proteins, including tissue factor (TF) and chemokine (C-C motif) ligand 2 (CCL2) in 2D-ADSC compared to that of the 3D-ADSC. Furthermore, 3D-ADSC containing 10,000 cells were reported to aggravate thrombosis and infarction in the livers [42], which indicates a necessity of control over spheroid size to protect the cells against hypoxia or nutrient deficiency after transplantation. The size-controlled MSC spheroids used for in vivo experiments in the current study survived well in a highly oxidative environment under in vitro conditions.…”

“…In a recent study, it was reported that the upregulation of CXCR4 expression in 3D aggregates promotes the adhesion of MSC spheroids with the endothelial cells [61], which would aid the localization of transplanted 3D spheroids at the site of injury. In another study, when bone marrow-derived mononuclear cell spheroids were delivered intraportally, they were found to be entrapped in the liver, whereas spheroid-derived single cells delivered intraportally drained into the systemic circulation [42].…”

Protective effect of a novel clinical-grade small molecule necrosis inhibitor against oxidative stress and inflammation during islet transplantation

“…Sections (4 mm) were stained using hematoxylin and eosin (H&E). Immunohistochemical staining were performed as previously described 30 . Briefly, after deparaffinization, 4‐μm tissue sections were stained with the polyclonal guinea pig anti‐insulin antibody (1:1000, A0546; Dako, Glostrup, Denmark) and mounted after counterstaining with 4ʹ‐6‐diamidino‐2‐phenylindole (DAPI).…”

Automation, Monitoring, and Standardization of Cell Product Manufacturing

“…In diabetes field, cell spheroids can provide an inducive environment for islet differentiation from stem cells, upregulate stemness factors and allow production of angiogenic and non-thrombogenic therapies ( Moritz et al, 2002 ; Oh et al, 2018 ; Lo et al, 2019 ). Early data of islets/mesenchymal stem cell co-cultures in spheroids demonstrated improved islet long term viability, but not function ( Rawal et al, 2017 ).…”