Micro/nanoscale technologies for the development of hormone-expressing islet-like cell clusters
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Insulin-expressing islet-like cell clusters derived from precursor cells have significant potential in the treatment of type-I diabetes. Given that cluster size and uniformity are known to influence islet cell behavior, the ability to effectively control these parameters could find applications in the development of anti-diabetic therapies. In this work, we combined micro and nanofabrication techniques to build a biodegradable platform capable of supporting the formation of islet-like structures from pancreatic precursors. Soft lithography and electrospinning were used to create arrays of microwells (150–500 μm diameter) structurally interfaced with a porous sheet of micro/nanoscale polyblend fibers (∼0.5–10 μm in cross-sectional size), upon which human pancreatic ductal epithelial cells anchored and assembled into insulin-expressing 3D clusters. The microwells effectively regulated the spatial distribution of the cells on the platform, as well as cluster size, shape and homogeneity. Average cluster cross-sectional area (∼14000–17500 μm2) varied in proportion to the microwell dimensions, and mean circularity values remained above 0.7 for all microwell sizes. In comparison, clustering on control surfaces (fibers without microwells or tissue culture plastic) resulted in irregularly shaped/sized cell aggregates. Immunoreactivity for insulin, C-peptide and glucagon was detected on both the platform and control surfaces; however, intracellular levels of C-peptide/cell were ∼60 % higher on the platform.
KeywordsDiabetes Soft lithography Electrospinning Precursor cell
The authors would like to thank the staff of the Ohio Nanotech West Laboratory for technical assistance, and Dr. Herbert Bresler for his helpful suggestions during the early phases of the work. Human islet-derived precursor cells were kindly donated by Dr. Gershengorn’s lab (NIH). This work was supported with funds from the Juvenile Diabetes Research Foundation (Grant No. 5-2009-511), the National Science Foundation (Grants No. EEC-0425626 and No. CMMI-0928739), and AFOSR MURI (Grant No. F49620-03-1-0421).
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