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.
The Islet Research Laboratory, Worcester Clinical Research Unit, Worcestershire Acute Hospitals NHS Trust, Worcester, UK.
Khalid M
KM
Qureshi
Jou
J
Lee
Michelle B
MB
Paget
Clifford J
CJ
Bailey
S John
SJ
Curnow
Hilary E
HE
Murray
Richard
R
Downing
D000328:Adult; D000650:Amnion / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000276:immunology; D005260:Female; D006801:Humans; D018472:Hypogravity; D056747:Immunomodulation; D007515:Islets of Langerhans / Q000276:immunology*; D016381:Islets of Langerhans Transplantation / Q000276:immunology / Q000379:methods*; D012399:Rotation; D013234:Stem Cells / Q000276:immunology*; D014184:Transplantation, Homologous / Q000379:methods
T cell; allo-reactivity; amniotic epithelial cells; immunomodulation; islet transplantation; rotational cell culture