Reproducing diabetic retinopathy features using newly developed human induced-pluripotent stem cell-derived retinal Müller glial cells.
Reproducing diabetic retinopathy features using newly developed human induced-pluripotent stem cell-derived retinal Müller glial cells.
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作者：
Couturier A;Blot G;Vignaud L;Nanteau C;Slembrouck-Brec A;Fradot V;Acar N;Sahel JA;Tadayoni R;Thuret G;Sennlaub F;Roger JE;Goureau O;Guillonneau X;Reichman S
Muller胶质细胞（MGCs）负责视网膜的稳态和代谢支持。尽管MGC在视网膜疾病中的重要性，但可靠和可获得的用于MGC相关疾病建模的人类细胞来源尚缺乏。虽然可以从死后视网膜组织中纯化原代人MGCs，但供体的稀缺性限制了其使用。为了克服这一问题，我们开发了一种方案来生成和储存人类诱导多能干细胞衍生的MGCs （hiMGCs）。通过转录组分析，我们发现生成的三种遗传独立的hiMGCs是均匀的，并显示出pMGCs的表型特征和转录组谱。这些细胞表达关键的MGC标记物，包括Vimentin、CLU、DKK3、SOX9、SOX2、S100A16、ITGB1和CD44，可以培养到8代。在我们的培养条件下，hiMGCs和pMGCs表达低水平的RLPB1、AQP4、KCNJ1、kcnj10和SLC1A3。通过疾病建模方法，我们发现hiMGCs可用于模拟糖尿病视网膜病变（DR）相关血脂异常的特征。事实上，棕榈酸盐是糖尿病患者血浆水平升高的一种主要游离脂肪酸，可诱导DR患者眼液中炎性细胞因子的表达，如CXCL8 （IL‐8）和ANGPTL4。此外，棕榈酸处理的hiMGC分泌组分析显示，与DR密切相关的促血管生成因子上调，包括ANG2、Endoglin、IL‐1β、CXCL8、MMP‐9、PDGF‐AA和VEGF。因此，hiMGCs可能是pMGCs的替代品，也是一种非常有价值的工具，可以帮助理解和模拟神经胶质细胞参与视网膜疾病，包括dr。我们开发了一种方案来生成和储存人类iPSC衍生的<s:1> ller神经胶质细胞（hiMGCs）。hiMGCs具有原发性MGCs的表型特征和转录组学特征。hiMGCs可以用来模拟糖尿病视网膜病变的特征。
Muller glial cells (MGCs) are responsible for the homeostatic and metabolic support of the retina. Despite the importance of MGCs in retinal disorders, reliable and accessible human cell sources to be used to model MGC‐associated diseases are lacking. Although primary human MGCs (pMGCs) can be purified from post‐mortem retinal tissues, the donor scarcity limits their use. To overcome this problem, we developed a protocol to generate and bank human induced pluripotent stem cell‐derived MGCs (hiMGCs). Using a transcriptome analysis, we showed that the three genetically independent hiMGCs generated were homogeneous and showed phenotypic characteristics and transcriptomic profile of pMGCs. These cells expressed key MGC markers, including Vimentin, CLU, DKK3, SOX9, SOX2, S100A16, ITGB1, and CD44 and could be cultured up to passage 8. Under our culture conditions, hiMGCs and pMGCs expressed low transcript levels of RLPB1, AQP4, KCNJ1, KCJN10, and SLC1A3. Using a disease modeling approach, we showed that hiMGCs could be used to model the features of diabetic retinopathy (DR)‐associated dyslipidemia. Indeed, palmitate, a major free fatty acid with elevated plasma levels in diabetic patients, induced the expression of inflammatory cytokines found in the ocular fluid of DR patients such as CXCL8 (IL‐8) and ANGPTL4. Moreover, the analysis of palmitate‐treated hiMGC secretome showed an upregulation of proangiogenic factors strongly related to DR, including ANG2, Endoglin, IL‐1β, CXCL8, MMP‐9, PDGF‐AA, and VEGF. Thus, hiMGCs could be an alternative to pMGCs and an extremely valuable tool to help to understand and model glial cell involvement in retinal disorders, including DR. We developed a protocol to generate and bank human iPSC‐derived Müller Glial cells (hiMGCs). hiMGCs showed phenotypic characteristics and transcriptomic profile of primary MGCs. hiMGCs can be used to model the features of diabetic retinopathy.
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DOI：
10.1056/nejmoa1001288
发表时间：
2010-07-15
期刊：
The New England journal of medicine
影响因子：
--
作者：
ACCORD Study Group;ACCORD Eye Study Group;Chew EY;Ambrosius WT;Davis MD;Danis RP;Gangaputra S;Greven CM;Hubbard L;Esser BA;Lovato JF;Perdue LH;Goff DC Jr;Cushman WC;Ginsberg HN;Elam MB;Genuth S;Gerstein HC;Schubart U;Fine LJ
通讯作者：
Fine LJ
影响因子：
1.8
作者：
Hammer SS;Busik JV
通讯作者：
Busik JV
DOI：
10.1097/iae.0000000000000109
发表时间：
2014-08-01
影响因子：
3.3
作者：
Jeon, Sohee;Lee, Won Ki
通讯作者：
Lee, Won Ki
影响因子：
168.9
作者：
Keech, A. C.;Mitchell, P.;Colman, P. G.
通讯作者：
Colman, P. G.
Müller细胞 - 微神经群横断性驱动糖尿病性视网膜病中的神经炎症。
影响因子：
7.7
作者：
Abcouwer SF
通讯作者：
Abcouwer SF