A culture substratum with net-like polyamide fibers promotes the differentiation of mouse and human pluripotent stem cells to insulin-producing cells
A culture substratum with net-like polyamide fibers promotes the differentiation of mouse and human pluripotent stem cells to insulin-producing cells
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网状聚酰胺纤维培养基促进小鼠和人类多能干细胞向胰岛素产生细胞的分化
DOI：
10.1088/1748-605x/ab261c
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发表时间：
2019
影响因子：
4
通讯作者：
Tomizawa Kazuhito
中科院分区：
文献类型：
--
作者：
Kaitsuka Taku;Kojima Rie;Kawabe Masaaki;Noguchi Hirofumi;Shiraki Nobuaki;Kume Shoen;Tomizawa Kazuhito
来源于小鼠多能干细胞（PSC）的胰岛素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。在此之前，我们已经建立了一个分化协议，从小鼠胚胎干细胞（ESC）获得胰岛素分泌细胞使用的生长因子，重组蛋白，和培养基质与网状纤维的组合。然而，还没有测试这些纤维的哪些材料和直径对于分化更有效。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质。根据纤维直径将基质划分为PA 100、300、600、PAN 100、300和600组。然后通过胰岛素1（Ins 1）表达检测在基质上培养的小鼠ESCs的分化效率。发现在PA 300分化细胞中表达最高，表明产生高水平胰岛素的潜力。为了了解基质性质的任何差异，测量层粘连蛋白的吸附能力，显示PA 300对层粘连蛋白的吸附能力最高。我们接下来检查了与PA 300孵育影响的分化阶段。这表明Sox 17-和Pdx 1-GFP-阳性细胞在分化的第一步中增加。为了显示胰岛素的产生而不从培养基中吸收，我们证实了分化后胰岛素C肽的表达。最后，我们测试了PA 300对人诱导的PSC分化的影响，并且发现在定形内胚层阶段具有PA 300基质的Sox 17阳性细胞更多。此外，这些细胞表达胰岛素C肽并具有葡萄糖响应性C肽分泌。总之，我们的研究鉴定并验证了一种适用于小鼠和人PSC胰腺分化的新型基质。
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. We next 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 glucose-responsive C-peptide secretion. In summary, our study identified and validated a novel substratum which is suitable for pancreatic differentiation of mouse and human PSCs.
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影响因子：
4
作者：
Higuchi, Yuichiro;Shiraki, Nobuaki;Kume, Shoen
通讯作者：
Kume, Shoen
影响因子：
3.5
作者：
Abazari, Mohammad Foad;Soleimanifar, Fatemeh;Kehtari, Mousa
通讯作者：
Kehtari, Mousa
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Tanabe, Koji;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya
影响因子：
5.2
作者：
Shiraki, Nobuaki;Yoshida, Tetsu;Kume, Shoen
通讯作者：
Kume, Shoen
VMAT2 被确定为晚期 β 细胞分化的调节因子。
DOI：
--
发表时间：
2014
期刊：
Nat. Chem. Biol.
影响因子：
--
作者：
Sakano;D.;Shiraki;N.;Kikawa;K.;Yamazoe;T.;Kataoka;M.;Umeda;K.;Araki;K.;Mao;D.;Matsumoto;S.;Nakagata;N.;Anderson;O.;Stainier;D.;Endo;F.;Kume;K.;Uesugi;M. and Kume;S.
通讯作者：
S.