Johansson Ulrika, Shalaly Nancy Dekki, Hjelm Linnea Charlotta, Ria Massimiliano, Berggren Per-Olof, Hedhammar My
Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, SE-106 91 Stockholm, Sweden.
Linnæus Center of Biomaterials Chemistry, Linnæus University, Kalmar, Sweden.
ACS Biomater Sci Eng. 2020 Feb 10;6(2):1186-1195. doi: 10.1021/acsbiomaterials.9b01573. Epub 2020 Jan 8.
Pancreatic islet transplantation has not yet succeeded as an overall treatment for type 1 diabetes because of limited access to donor islets, as well as low efficacy and poor reproducibility of the current procedure. Herein, a method to create islets-like composite clusters (coclusters) from dispersed endocrine cells and supportive cells is described, attempting to improve compatibility with the recipient and more efficiently make use of the donor-derived material. To mimic the extracellular matrix environment, recombinant spider silk functionalized with cell binding motifs are used as 3D support for the coclusters. A cell binding motif derived from fibronectin (FN) was found superior in promoting cell adherence, while a plain RGD-motif incorporated in the repetitive part of the silk protein (2R) increased the mobility and cluster formation of endocrine cells. Self-assembly of a mixture of FN/2R silk is utilized to integrate endocrine cells together with endothelial and mesenchymal cells into islet-like coclusters. Both xenogenic and allogenic versions of these coclusters were found to be viable and were able to respond to dynamic glucose stimulation with insulin release. Moreover, the endothelial cells were found to be colocalized with the endocrine cells, showing that the silk combined with supportive cells may promote vascularization. This method to engineer combined islet-like coclusters allows donor-derived endocrine cells to be surrounded by supportive cells from the recipient, which have the potential to further promote engraftment in the host and considerably reduce risk of rejection.
由于供体胰岛获取有限，以及当前手术的疗效低和可重复性差，胰岛移植尚未成功成为1型糖尿病的整体治疗方法。本文描述了一种从分散的内分泌细胞和支持细胞创建胰岛样复合簇（共簇）的方法，试图提高与受体的相容性，并更有效地利用供体来源的材料。为了模拟细胞外基质环境，用细胞结合基序功能化的重组蜘蛛丝被用作共簇的三维支持物。发现源自纤连蛋白（FN）的细胞结合基序在促进细胞黏附方面更具优势，而掺入丝蛋白重复部分的普通RGD基序（2R）增加了内分泌细胞的迁移率和簇形成。利用FN/2R丝混合物的自组装将内分泌细胞与内皮细胞和间充质细胞整合到胰岛样共簇中。发现这些共簇的异种和同种异体版本都是有活力的，并且能够通过胰岛素释放对动态葡萄糖刺激作出反应。此外，发现内皮细胞与内分泌细胞共定位，表明丝与支持细胞结合可能促进血管生成。这种构建联合胰岛样共簇的方法使供体来源的内分泌细胞被受体的支持细胞包围，这有可能进一步促进在宿主中的植入，并大大降低排斥风险。
