Microchip-based engineering of super-pancreatic islets supported by adipose-derived stem cells.

Type 1 diabetes mellitus (T1DM) is a chronic disorder characterized by targeted autoimmune-mediated destruction of the β cells of Langerhans within pancreatic islets. Currently, islet transplantation is the only curative therapy; however, donor shortages and cellular damage during the isolation process critically limit the use of this approach. Here, we describe a method for creating viable and functionally potent islets for successful transplantation by co-culturing single primary islet cells with adipose-derived stem cells (ADSCs) in concave microwells. We observed that the ADSCs segregated from the islet cells, eventually yielding purified islet spheroids in the three-dimensional environment. Thereafter, the ADSC-exposed islet spheroids showed significantly different ultrastructural morphologies, higher viability, and enhanced insulin secretion compared to mono-cultured islet spheroids. This suggests that ADSCs may have a significant potential to protect islet cells from damage during culture, and may be employed to improve islet cell survival and function prior to transplantation. In vivo experiments involving xenotransplantation of microfiber-encapsulated spheroids into a mouse model of diabetes revealed that co-culture-transplanted mice maintained their blood glucose levels longer than mono-culture-transplanted mice, and required less islet mass to reverse diabetes. This method for culturing islet spheroids could potentially help overcome the cell shortages that have limited clinical applications and could possibly be developed into a bioartificial pancreas.

[1]  Peter M. Jones,et al.  Pre-culturing islets with mesenchymal stromal cells using a direct contact configuration is beneficial for transplantation outcome in diabetic mice. , 2013, Cytotherapy.

[2]  D. Kaplan,et al.  Enhanced function of pancreatic islets co-encapsulated with ECM proteins and mesenchymal stromal cells in a silk hydrogel. , 2012, Biomaterials.

[3]  S. Paraskevas,et al.  Cell Loss in Isolated Human Islets Occurs by Apoptosis , 2000, Pancreas.

[4]  Bong Geun Chung,et al.  Concave microwell based size-controllable hepatosphere as a three-dimensional liver tissue model. , 2011, Biomaterials.

[5]  Dong Yun Lee,et al.  In situ formation and collagen-alginate composite encapsulation of pancreatic islet spheroids. , 2012, Biomaterials.

[6]  K. Lee,et al.  Diffusion-mediated in situ alginate encapsulation of cell spheroids using microscale concave well and nanoporous membrane. , 2011, Lab on a chip.

[7]  Min Jun Kim,et al.  Mutual effect of subcutaneously transplanted human adipose-derived stem cells and pancreatic islets within fibrin gel. , 2013, Biomaterials.

[8]  M. Grompe,et al.  Generation and Regeneration of Cells of the Liver and Pancreas , 2008, Science.

[9]  G. Cooper,et al.  ULTRASTRUCTURAL EVIDENCE THAT APOPTOSIS IS THE MECHANISM BY WHICH HUMAN AMYLIN EVOKES DEATH IN RINm5F PANCREATIC ISLET β‐CELLS , 2001, Cell biology international.

[10]  Min Jun Kim,et al.  Functional clustering of pancreatic islet cells using concave microwell array , 2011 .

[11]  T. Utsunomiya,et al.  Human adipose-derived stem cells: Potential clinical applications in surgery , 2010, Surgery Today.

[12]  Song-Cheol Kim,et al.  Bone marrow-derived mesenchymal stromal cells support rat pancreatic islet survival and insulin secretory function in vitro. , 2011, Cytotherapy.

[13]  Sang-Hoon Lee,et al.  Cell encapsulation via microtechnologies. , 2014, Biomaterials.

[14]  J. Schrezenmeir,et al.  Effect of the immunosuppressive regime of Edmonton protocol on the long-term in vitro insulin secretion from islets of two different species and age categories. , 2005, Toxicology in vitro : an international journal published in association with BIBRA.

[15]  S. Sumi,et al.  Encapsulated islets transplantation: Past, present and future. , 2012, World journal of gastrointestinal pathophysiology.

[16]  L. Buscail,et al.  Adult Stromal Cells Derived from Human Adipose Tissue Provoke Pancreatic Cancer Cell Death both In Vitro and In Vivo , 2009, PloS one.

[17]  M. Kassem,et al.  Human bone-marrow-derived mesenchymal stem cells: biological characteristics and potential role in therapy of degenerative diseases , 2007, Cell and Tissue Research.

[18]  P. Seglen.  Preparation of isolated rat liver cells. , 1976, Methods in cell biology.

[19]  A. Monaco,et al.  An improved method for isolation of mouse pancreatic islets. , 1985, Transplantation.

[20]  Han-Hung Huang,et al.  A replacement for islet equivalents with improved reliability and validity , 2012, Acta Diabetologica.

[21]  Marília Mateus,et al.  An Overview on the Development of a Bio‐Artificial Pancreas as a Treatment of Insulin‐Dependent Diabetes Mellitus , 2006 .

[22]  G. Michalopoulos,et al.  Liver Regeneration , 1997, Science.

[23]  Ali Khademhosseini,et al.  Controlled-size embryoid body formation in concave microwell arrays. , 2010, Biomaterials.

[24]  J. Permert,et al.  Optimising islet engraftment is critical for successful clinical islet transplantation , 2008, Diabetologia.

[25]  L. Stehno-Bittel,et al.  Small rat islets are superior to large islets in in vitro function and in transplantation outcomes. , 2006, American journal of physiology. Endocrinology and metabolism.

[26]  Y. Bae,et al.  Human Adipose Tissue-Derived Mesenchymal Stem Cells Improve Postnatal Neovascularization in a Mouse Model of Hindlimb Ischemia , 2006, Cellular Physiology and Biochemistry.

[27]  G. Duruksu,et al.  Protection of rat pancreatic islet function and viability by coculture with rat bone marrow-derived mesenchymal stem cells , 2010, Cell Death and Disease.

[28]  J. Gimble,et al.  Adipose-derived stem cells for regenerative medicine. , 2007, Circulation research.

[29]  George K Michalopoulos,et al.  Liver regeneration. , 2005, Advances in biochemical engineering/biotechnology.

[30]  L. Stehno-Bittel,et al.  Engineering islets for improved performance by optimized reaggregation in a micromold. , 2013, Tissue engineering. Part A.

[31]  N. Barshes,et al.  islet transplantation: implications for intrahepatic grafts , 2022 .

[32]  Wolfgang Moritz,et al.  Superiority of Small Islets in Human Islet Transplantation , 2007, Diabetes.

[33]  M. Corbascio,et al.  Small Islets are Essential for Successful Intraportal Transplantation in a Diabetes Mouse Model , 2010, Scandinavian journal of immunology.

[34]  S. Arden,et al.  The post-translational processing and intracellular sorting of carboxypeptidase H in the islets of Langerhans , 1995, Molecular and Cellular Endocrinology.

[35]  J. Stockman,et al.  International Trial of the Edmonton Protocol for Islet Transplantation , 2008 .

[36]  T. Lohmann,et al.  Diabetes mellitus and islet cell specific autoimmunity as adverse effects of immunsuppressive therapy by FK506/tacrolimus. , 2000, Experimental and clinical endocrinology & diabetes : official journal, German Society of Endocrinology [and] German Diabetes Association.

[37]  Christophe Benoist,et al.  β-Cell death during progression to diabetes , 2001, Nature.

[38]  C. Stabler,et al.  Enhancing Clinical Islet Transplantation through Tissue Engineeering Strategies , 2010, Journal of diabetes science and technology.

[39]  Min Zhu,et al.  Comparison of Multi-Lineage Cells from Human Adipose Tissue and Bone Marrow , 2003, Cells Tissues Organs.

[40]  P. Arner,et al.  Functional studies of mesenchymal stem cells derived from adult human adipose tissue. , 2005, Experimental cell research.

[41]  S. Messinger,et al.  Improved Human Islet Isolation Using Nicotinamide , 2006, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[42]  S. Bonner-Weir,et al.  Spontaneous Reassociation of Dispersed Adult Rat Pancreatic Islet Cells Into Aggregates With Three-Dimensional Architecture Typical of Native Islets , 1987, Diabetes.

[43]  U. Boggi,et al.  Insulin secretory function is impaired in isolated human islets carrying the Gly(972)-->Arg IRS-1 polymorphism. , 2002, Diabetes.

[44]  P. Squires,et al.  Pancreatic beta-cell-to-beta-cell interactions are required for integrated responses to nutrient stimuli: enhanced Ca2+ and insulin secretory responses of MIN6 pseudoislets. , 1999, Diabetes.

[45]  Shing-Hwa Liu,et al.  A Novel Human Stem Cell Coculture System that Maintains the Survival and Function of Culture Islet-Like Cell Clusters , 2008, Cell transplantation.

[46]  Sang-Hoon Lee,et al.  Microfluidic spinning of micro- and nano-scale fibers for tissue engineering. , 2014, Lab on a chip.

[47]  D. Melton,et al.  Organ size is limited by the number of embryonic progenitor cells in the pancreas but not the liver , 2007, Nature.

[48]  C. Benoist,et al.  b-Cell death during progression to diabetes , 2001 .

[49]  Sang-Hoon Lee,et al.  Size-controllable networked neurospheres as a 3D neuronal tissue model for Alzheimer's disease studies. , 2013, Biomaterials.

[50]  Y. Takeda,et al.  Combined Transplantation of Pancreatic Islets and Adipose Tissue-Derived Stem Cells Enhances the Survival and Insulin Function of Islet Grafts in Diabetic Mice , 2010, Transplantation.

[51]  Dong Yun Lee,et al.  3D co-culturing model of primary pancreatic islets and hepatocytes in hybrid spheroid to overcome pancreatic cell shortage. , 2013, Biomaterials.

[52]  Keith L. March,et al.  Secretion of Angiogenic and Antiapoptotic Factors by Human Adipose Stromal Cells , 2004, Circulation.

[53]  Min Jun Kim,et al.  Microfluidics-generated pancreatic islet microfibers for enhanced immunoprotection. , 2013, Biomaterials.

[54]  Yin-ping Li,et al.  Adipose-derived mesenchymal stem cells ameliorate STZ-induced pancreas damage in type 1 diabetes. , 2012, Bio-medical materials and engineering.

[55]  DoYeun Park,et al.  Functional 3D Human Primary Hepatocyte Spheroids Made by Co-Culturing Hepatocytes from Partial Hepatectomy Specimens and Human Adipose-Derived Stem Cells , 2012, PloS one.