Improved islet recovery and efficacy through co-culture and co-transplantation of islets with human adipose-derived mesenchymal stem cells

Abstract

Islet transplantation is an established clinical procedure for select patients with type 1 diabetes and severe hypoglycemia to stabilize glycemic control. Post-transplant, substantial beta cell mass is lost, necessitating multiple donors to maintain euglycemia. A potential strategy to augment islet engraftment is the co-transplantation of islets with multipotent mesenchymal stem cells to capitalize upon their pro-angiogenic and anti-inflammatory properties. Herein, we examine the in vitro and in vivo effect of co-culturing murine islets with human adipose-derived mesenchymal stem cells (Ad-MSCs). Islets co-cultured with Ad-MSCs for 48 hours had decreased cell death, superior viability as measured by membrane integrity, improved glucose stimulated insulin secretion and reduced apoptosis compared to control islets. These observations were recapitulated with human islets, albeit tested in a limited capacity. Recipients of marginal mouse islet mass grafts, co-transplanted with Ad-MSCs without a co-culture period, did not reverse to normoglycemia as efficiently as islets alone. However, utilizing a 48-hour co-culture period, marginal mouse islets grafts with Ad-MSCs achieved a superior percent euglycemia rate when compared to islets cultured and transplanted alone. A co-culture period of human islets with human Ad-MSCs may have a clinical benefit improving engraftment outcomes.

Amniotic Membrane Extract Protects Islets From Serum-Deprivation Induced Impairments and Improves Islet Transplantation Outcome

“…To determine islet yield, islets prior to and post 48-h culture were harvested and counted as previously described ( 36 ). Briefly, aliquots from both experimental and control groups were stained with dithizone (Sigma Aldrich, USA) and counted.…”

“…Apoptosis of the cultured islets was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay (TUNEL)staining and flow cytometry as previously described (36)(37)(38). For TUNEL staining analysis, briefly, islets were collected and fixed in 4% paraformaldehyde, embedded in 3% agar (A8190, Solarbio) to create a solid cellularmatrix interaction, then processed and embedded in paraffin.…”

Bioprocess Development for Human Mesenchymal Stem Cell Therapy Products

“…In both applications, large numbers of functional beta cells are required, but beta cells rapidly lose their functionality when expanded in vitro. The loss of beta cell functionality in vitro can be prevented by cocultivation with MSCs, which not only stimulate beta cell proliferation but also enhance their glucose-dependent secretion of insulin [97][98][99].…”

Exosomes derived from human mesenchymal stem cells preserve mouse islet survival and insulin secretion function

“…Several studies have confirmed the positive effect of MSCs on islet viability and function through paracrine interactions and/or direct contact both in the culture period and after transplantation (Rackham et al, 2014[ 29 ]; Scuteri et al, 2014[ 35 ]; Yamada et al, 2014[ 42 ]). Some report that the protective effect of MSCs on cultured islets occurs via direct contact and the production of extracellular matrix molecules and annexin A1 (Arzouni et al, 2017[ 2 ]; Gamble et al, 2018[ 11 ]; Montanari et al, 2017[ 24 ]). This is while others suggest that indirect co-culture systems involving Transwell® inserts or conditioned media of MSCs (MSC-CM) could also inhibit apoptosis and promote islet viability and function through the secretion of trophic factors (de Souza et al, 2017[ 8 ]; Schive et al, 2017[ 34 ]; Yamada et al, 2014[ 42 ]).…”