Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal Changes
Pancreatic islet transplantation is the fundamental treatment for insulin-dependent diabetes; however, donor shortage is a major hurdle in its use as a standard treatment. Accordingly, differentiated insulin-producing cells (DIPCs) are being developed as a new islet source. Differentiation efficienc...
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MDPI AG
2020-11-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/12/2551 |
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| author | Yu Na Lee Hye Jin Yi Hanse Goh Ji Yoon Park Sarah Ferber In Kyong Shim Song Cheol Kim |
| author_facet | Yu Na Lee Hye Jin Yi Hanse Goh Ji Yoon Park Sarah Ferber In Kyong Shim Song Cheol Kim |
| author_sort | Yu Na Lee |
| collection | DOAJ |
| description | Pancreatic islet transplantation is the fundamental treatment for insulin-dependent diabetes; however, donor shortage is a major hurdle in its use as a standard treatment. Accordingly, differentiated insulin-producing cells (DIPCs) are being developed as a new islet source. Differentiation efficiency could be enhanced if the spheroid structure of the natural islets could be recapitulated. Here, we fabricated DIPC spheroids using concave microwells, which enabled large-scale production of spheroids of the desired size. We prepared DIPCs from human liver cells by trans-differentiation using transcription factor gene transduction. Islet-related gene expression and insulin secretion levels were higher in spheroids compared to those in single-cell DIPCs, whereas actin–myosin interactions significantly decreased. We verified actin–myosin-dependent insulin expression in single-cell DIPCs by using actin–myosin interaction inhibitors. Upon transplanting cells into the kidney capsule of diabetic mouse, blood glucose levels decreased to 200 mg/dL in spheroid-transplanted mice but not in single cell-transplanted mice. Spheroid-transplanted mice showed high engraftment efficiency in in vivo fluorescence imaging. These results demonstrated that spheroids fabricated using concave microwells enhanced the engraftment and functions of DIPCs via actin–myosin-mediated cytoskeletal changes. Our strategy potentially extends the clinical application of DIPCs for improved differentiation, glycemic control, and transplantation efficiency of islets. |
| first_indexed | 2024-03-10T14:30:28Z |
| format | Article |
| id | doaj.art-8e65598dc10b473fa5a95f8153473ca5 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T14:30:28Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-8e65598dc10b473fa5a95f8153473ca52023-11-20T22:38:29ZengMDPI AGCells2073-44092020-11-01912255110.3390/cells9122551Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal ChangesYu Na Lee0Hye Jin Yi1Hanse Goh2Ji Yoon Park3Sarah Ferber4In Kyong Shim5Song Cheol Kim6Asan Medical Center, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Seoul 05505, KoreaAsan Medical Center, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Seoul 05505, KoreaAsan Medical Center, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Seoul 05505, KoreaAsan Medical Center, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Seoul 05505, KoreaRegenerative Medicine, Stem Cell and Tissue Engineering Center, Sheba Medical Center, Tel-Hashomer 52621, IsraelAsan Medical Center, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Seoul 05505, KoreaAsan Medical Center, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Seoul 05505, KoreaPancreatic islet transplantation is the fundamental treatment for insulin-dependent diabetes; however, donor shortage is a major hurdle in its use as a standard treatment. Accordingly, differentiated insulin-producing cells (DIPCs) are being developed as a new islet source. Differentiation efficiency could be enhanced if the spheroid structure of the natural islets could be recapitulated. Here, we fabricated DIPC spheroids using concave microwells, which enabled large-scale production of spheroids of the desired size. We prepared DIPCs from human liver cells by trans-differentiation using transcription factor gene transduction. Islet-related gene expression and insulin secretion levels were higher in spheroids compared to those in single-cell DIPCs, whereas actin–myosin interactions significantly decreased. We verified actin–myosin-dependent insulin expression in single-cell DIPCs by using actin–myosin interaction inhibitors. Upon transplanting cells into the kidney capsule of diabetic mouse, blood glucose levels decreased to 200 mg/dL in spheroid-transplanted mice but not in single cell-transplanted mice. Spheroid-transplanted mice showed high engraftment efficiency in in vivo fluorescence imaging. These results demonstrated that spheroids fabricated using concave microwells enhanced the engraftment and functions of DIPCs via actin–myosin-mediated cytoskeletal changes. Our strategy potentially extends the clinical application of DIPCs for improved differentiation, glycemic control, and transplantation efficiency of islets.https://www.mdpi.com/2073-4409/9/12/2551insulin-producing cellsspheroidthree-dimensional cultureconcave microwelldiabetescytoskeleton changes |
| spellingShingle | Yu Na Lee Hye Jin Yi Hanse Goh Ji Yoon Park Sarah Ferber In Kyong Shim Song Cheol Kim Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal Changes Cells insulin-producing cells spheroid three-dimensional culture concave microwell diabetes cytoskeleton changes |
| title | Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal Changes |
| title_full | Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal Changes |
| title_fullStr | Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal Changes |
| title_full_unstemmed | Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal Changes |
| title_short | Spheroid Fabrication Using Concave Microwells Enhances the Differentiation Efficacy and Function of Insulin-Producing Cells via Cytoskeletal Changes |
| title_sort | spheroid fabrication using concave microwells enhances the differentiation efficacy and function of insulin producing cells via cytoskeletal changes |
| topic | insulin-producing cells spheroid three-dimensional culture concave microwell diabetes cytoskeleton changes |
| url | https://www.mdpi.com/2073-4409/9/12/2551 |
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