Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin.
Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin.
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DOI：
10.1167/iovs.17-23157
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发表时间：
2018-06-01
影响因子：
4.4
通讯作者：
Singh R
中科院分区：
文献类型：
--
作者：
Galloway CA;Dalvi S;Shadforth AMA;Suzuki S;Wilson M;Kuai D;Hashim A;MacDonald LA;Gamm DM;Harkin DG;Singh R
RPE细胞移植作为AMD的潜在治疗已经被广泛研究;然而，在AMD中，超微结构损伤影响RPE及其底层基质支持物布鲁赫膜（Bruch’s membrane，BrM）。因此，由替代支架支撑的RPE单层可以为AMD的基于细胞的治疗提供更有效的方法。为了实现这一目标，我们的目的是建立一个功能性的人诱导多能干细胞衍生（hiPSC）-RPE单层的桑蚕丝素蛋白（BMSF）支架。将从五种不同的hiPSC系分化的RPE在涂覆有细胞外基质（ECM，COL 1）的BMSF膜上培养，并且在常规组织培养塑料或涂覆有ECM（LAM-TCP）的Transwell上培养。在1天至≥3个月的纵向实验中，比较了在不同膜上培养的hiPSC-RPE的形态、基因和蛋白表达以及功能特征。ECM包被的BMSF和TCP上的hiPSC-RPE单层可以在培养中维持≥3个月，并显示RPE特征性形态、色素沉着、极性以及RPE标签基因和蛋白的表达。此外，ECM包被的BMSF和TCP上的hiPSC-RPE显示出几种基底膜蛋白的稳健表达和分泌。重要的是，COL 1-BMSF和LAM-TCP上的hiPSC-RPE细胞在光感受器外节的吞噬和降解中显示出相似的功效。由丝素蛋白制成的生物材料支架支持功能性hiPSC-RPE单层的成熟和长期存活。这对体外疾病建模和体内细胞替代疗法都具有重要意义。
RPE cell transplantation as a potential treatment for AMD has been extensively investigated; however, in AMD, ultrastructural damage affects both the RPE and its underlying matrix support, the Bruch's membrane (BrM). An RPE monolayer supported by a surrogate scaffold could thus provide a more effective approach to cell-based therapy for AMD. Toward this goal, we aimed to establish a functional human induced pluripotent stem cell–derived (hiPSC)-RPE monolayer on a Bombyx mori silk fibroin (BMSF) scaffold. RPE differentiated from five distinct hiPSC lines were cultured on BMSF membrane coated with extracellular matrix (ECM, COL1), and either regular tissue culture plastic or Transwell coated with ECM (LAM-TCP). Morphologic, gene and protein expression, and functional characteristics of the hiPSC-RPE cultured on different membranes were compared in longitudinal experiments spanning 1 day to ≥3 months. The hiPSC-RPE monolayers on ECM-coated BMSF and TCP could be maintained in culture for ≥3 months and displayed RPE-characteristic morphology, pigmentation, polarity, and expression of RPE signature genes and proteins. Furthermore, hiPSC-RPE on both ECM-coated BMSF and TCP displayed robust expression and secretion of several basement membrane proteins. Importantly, hiPSC-RPE cells on COL1-BMSF and LAM-TCP showed similar efficacy in the phagocytosis and degradation of photoreceptor outer segments. A biomaterial scaffold manufactured from silk fibroin supports the maturation and long-term survival of a functional hiPSC-RPE monolayer. This has significant implications for both in vitro disease modeling and in vivo cell replacement therapy.
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影响因子：
41.2
作者：
Maya-Vetencourt JF;Ghezzi D;Antognazza MR;Colombo E;Mete M;Feyen P;Desii A;Buschiazzo A;Di Paolo M;Di Marco S;Ticconi F;Emionite L;Shmal D;Marini C;Donelli I;Freddi G;Maccarone R;Bisti S;Sambuceti G;Pertile G;Lanzani G;Benfenati F
通讯作者：
Benfenati F
影响因子：
4.4
作者：
Phillips, M. Joseph;Wallace, Kyle A.;Gamm, David M.
通讯作者：
Gamm, David M.
影响因子：
1.4
作者：
Jha, Balendu Shekhar;Bharti, Kapil
通讯作者：
Bharti, Kapil
影响因子：
17.8
作者：
Kur, Joanna;Newman, Eric A.;Chan-Ling, Tailoi
通讯作者：
Chan-Ling, Tailoi
影响因子：
3.3
作者：
Shadforth, Audra M. A.;Suzuki, Shuko;Harkin, Damien G.
通讯作者：
Harkin, Damien G.