Low gravity rotational culture and the integration of immunomodulatory stem cells reduce human islet allo-reactivity.
Modification of human islets prior to transplantation may improve long-term clinical outcome in terms of diabetes management, by supporting graft function and reducing the potential for allo-rejection. Intragraft incorporation of stem cells secreting beta (β)-cell trophic and immunomodulatory factors represents a credible approach, but requires suitable culture methods to facilitate islet alteration without compromising integrity. This study employed a three-dimensional rotational cell culture system (RCCS) to achieve modification, preserve function, and ultimately influence immune cell responsiveness to human islets. Islets underwent intentional dispersal and rotational culture-assisted aggregation with amniotic epithelial cells (AEC) exhibiting intrinsic immunomodulatory potential. Reassembled islet constructs were assessed for functional integrity, and their ability to induce an allo-response in discrete T-cell subsets determined using mixed islet:lymphocyte reaction assays. RCCS supported the formation of islet:AEC aggregates with improved insulin secretory capacity compared to unmodified islets. Further, the allo-response of peripheral blood mononuclear cell (PBMC) and purified CD4+ and CD8+ T-cell subsets to AEC-bearing grafts was significantly (p < 0.05) attenuated. Rotational culture enables pre-transplant islet modification involving their integration with immunomodulatory stem cells capable of subduing the allo-reactivity of T cells relevant to islet rejection. The approach may play a role in achieving acute and long-term graft survival in islet transplantation.
['Qureshi|Khalid M|KM|', 'Lee|Jou|J|', 'Paget|Michelle B|MB|', 'Bailey|Clifford J|CJ|', 'Curnow|S John|SJ|', 'Murray|Hilary E|HE|', 'Downing|Richard|R|']
D000328:Adult D000650:Amnion D018929:Cell Culture Techniques D002478:Cells, Cultured D004847:Epithelial Cells D005260:Female D006801:Humans D018472:Hypogravity D056747:Immunomodulation D007515:Islets of Langerhans D016381:Islets of Langerhans Transplantation D012399:Rotation D013234:Stem Cells D014184:Transplantation, Homologous
allo-reactivity rotational cell culture amniotic epithelial cells T cell islet transplantation immunomodulation