Enhanced Differentiation Capacity and Transplantation Efficacy of Insulin-Producing Cell Clusters from Human iPSCs Using Permeable Nanofibrous Microwell-Arrayed Membrane for Diabetes Treatment

Abstract

Although pancreatic islet transplantation is a potentially curative treatment for insulin-dependent diabetes, a shortage of donor sources, low differentiation capacity, and transplantation efficacy are major hurdles to overcome before becoming a standard therapy. Stem cell-derived insulin-producing cells (IPCs) are a potential approach to overcoming these limitations. To improve the differentiation capacity of the IPCs, cell cluster formation is crucial to mimic the 3D structure of the islet. This study developed a biodegradable polycaprolactone (PCL) electrospun nanofibrous (NF) microwell-arrayed membrane permeable to soluble factors. Based on the numerical analysis and experimental diffusion test, the NF microwell could provide sufficient nutrients, unlike an impermeable PDMS (polydimethylsiloxane) microwell. The IPC clusters in the NF microwells showed higher gene expression of insulin and PDX1 and insulin secretion than the PDMS microwells. The IPC clusters in the NF microwell-arrayed membrane could be directly transplanted. Transplanted IPC clusters in the microwells survived well and expressed PDX1 and insulin. Additionally, human c-peptide was identified in the blood plasma at two months after transplantation of the membranes. The NF microwell-arrayed membrane can be a new platform promoting IPC differentiation capacity and realizing an in situ transplantation technique for diabetic patients.

Successful Islet Transplantation Into a Subcutaneous Polycaprolactone Scaffold in Mice and Pigs

“…[21][22][23] Furthermore, it supports islet functional survival 21,22 and seems valuable for directing the differentiation of less mature insulin-producing cells. 24,25 In addition, the properties of PCL allow relatively easy addition of amino acids, growth factors, or immunomodulatory components. [26][27][28] The transition from rodent models to large preclinical models is always challenging.…”

Recent Progress in Fabrication of Electrospun Nanofiber Membranes for Developing Physiological In Vitro Organ/Tissue Models

“…[142] Shim et al generated hiPSCderived insulin-producing spheroids in a thermoformed PCL NF microwell array, which showed enhanced differentiation capacity compared with the traditional impermeable PDMS microwell array. [101] Interestingly, in the studies of Kim et al, Leet et al, and Shim et al, they integrated the 2.5D NFM composed of NF microwells with a cell culture insert, which not only provided a user-friendly cell culture platform but also supplied enough oxygen and nutrients/growth factors to spheroids from both the apical and basolateral sides of the well plate.…”

Electrospun Materials for Biomedical Applications

“…Kim, D., Kim, S., and co-authors detailed in their article entitled Enhanced Differentiation Capacity and Transplantation Efficacy of Insulin-Producing Cell Clusters from Human iPSCs Using Permeable Nanofibrous Microwell-Arrayed Membrane for Diabetes Treatment [ 30 ] a biodegradable polycaprolactone (PCL) electrospun nanofibrous microwell arrayed membrane permeable to soluble factors to improve some challenges related to insulin-dependent diabetes therapy. Stem cell-derived insulin producing cells (IPCs) were seeded into the electrospun microwell with a great cell survival rate.…”