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首页> 外文期刊>Biomedical materials >A culture substratum with net-like polyamide fibers promotes the differentiation of mouse and human pluripotent stem cells to insulinproducing cells
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A culture substratum with net-like polyamide fibers promotes the differentiation of mouse and human pluripotent stem cells to insulinproducing cells

机译：具有网状聚酰胺纤维的培养基促进小鼠和人类多能干细胞的分化为胰岛素激素的细胞

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摘要

Insulin-producing and -secreting cells derived from mouse pluripotent stem cells (PSCs) are useful for pancreatic development research and evaluating drugs that may induce insulin secretion. Previously, we have established a differentiation protocol to derive insulin-secreting cells from mouse embryonic stem cells (ESCs) using a combination of growth factors, recombinant proteins, and a culture substratum with net-like fibers. However, it has not been tested which materials and diameters of these fibers are more effective for the differentiation. Therefore, the present study aimed to produce net-like culture substratum formed from polyamide (PA) and polyacrylonitrile (PAN) fibers. Substrata were delineated into PA100, 300, 600, PAN100, 300, and 600 groups based on fiber diameters. The differentiation efficiencies of mouse ESCs cultured on the substrata were then examined by insulin 1 (Ins1) expression. Expression was found to be highest in PA300 differentiated cells, indicating the potential to produce high levels of insulin. To understand any differences in substratum properties, the adsorption capacities of laminin were measured, revealing that PA300 had the highest for it.Wenext examined the stage of differentiation affected by incubation with PA300. This showed that Sox17- and Pdx1-GFP-positive cells increased during the first step of differentiation. To show the production of insulin without absorption from the medium, we confirmed the expression of insulin C-peptide after differentiation. Finally, we tested the effects of PA300 on the differentiation of human-induced PSC, and found more Sox17-positive cells with the PA300 substratum at the definitive endoderm stage. Furthermore, these cells expressed insulin C-peptide and had glucoseresponsive C-peptide secretion. In summary, our study identified and validated a novel substratum which is suitable for pancreatic differentiation of mouse and human PSCs.
机译：衍生自小鼠多能干细胞（PSC）的胰岛素的产生和 - 分泌细胞可用于胰腺发育研究和评估可能诱导胰岛素分泌的药物。以前，我们已经建立了使用生长因子，重组蛋白质和培养基与网状纤维的培养基和培养基中的小鼠胚胎干细胞（ESC）从小鼠胚胎干细胞（ESC）中衍生胰岛素分泌细胞。然而，尚未测试这些纤维的材料和直径对分化更有效。因此，本研究旨在生产由聚酰胺（PA）和聚丙烯腈（PAN）纤维形成的网状培养基。基于纤维直径将SubStata描绘成PA100,300,600，Pan100,300和600组。然后通过胰岛素1（INS1）表达检查在亚样品上培养的小鼠ESC的分化效率。发现表达在PA300分化细胞中是最高的，表明产生高水平胰岛素的可能性。为了了解底层特性的任何差异，测量了层粘连蛋白的吸附能力，揭示了PA300对其最高的.Wenext检查了通过与PA300孵育影响的分化阶段。这表明SOX17-和PDX1-GFP阳性细胞在分化的第一步期间增加。为了显示不吸收培养基的胰岛素的生产，我们证实了分化后胰岛素C-肽的表达。最后，我们测试了PA300对人诱导的PSC分化的影响，并在最终的内胚层阶段在PA300癌中发现了更多SOx17阳性细胞。此外，这些细胞表达了胰岛素C-肽并具有葡糖蛋白酶C-肽分泌。总之，我们的研究鉴定并验证了一种新型底层，适用于小鼠和人PSC的胰腺分化。

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  • 来源
    《Biomedical materials》 |2019年第4期|共10页
  • 作者

    Taku Kaitsuka; Rie Kojima; Masaaki Kawabe; Hirofumi Noguchi; Nobuaki Shiraki; Shoen Kume; Kazuhito Tomizawa;

  • 作者单位

    Department of Molecular Physiology Faculty of Life Sciences Kumamoto University 1-1-1 Honjo Chuo-ku Kumamoto 860-8556 Japan;

    Central Research Laboratory Japan Vilene Company Ltd 7 Kitatone Koga 306-0213 Japan;

    Central Research Laboratory Japan Vilene Company Ltd 7 Kitatone Koga 306-0213 Japan;

    Department of Regenerative Medicine Graduate School of Medicine University of the Ryukyus Nishihara-cho Okinawa 903-0215 Japan;

    Department of Life Science and Technology School of Life Science and Technology Tokyo Institute of Technology 4259-B-25 Nagatsuta-cho Midori-ku Yokohama 226-8501 Japan;

    Department of Life Science and Technology School of Life Science and Technology Tokyo Institute of Technology 4259-B-25 Nagatsuta-cho Midori-ku Yokohama 226-8501 Japan;

    Department of Molecular Physiology Faculty of Life Sciences Kumamoto University 1-1-1 Honjo Chuo-ku Kumamoto 860-8556 Japan;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 医用一般科学;
  • 关键词

    pancreatic differentiation; embryonic stem cell; induced pluripotent stem cell; culture substratum; polyamide fiber; laminin;

    机译：胰腺分化;胚胎干细胞;诱导多能干细胞;培养基;聚酰胺纤维;薄片;

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