来源于人胚胎干细胞的三维早期视网膜祖细胞 3D 组织构建体。
Three-dimensional early retinal progenitor 3D tissue constructs derived from human embryonic stem cells.
机构信息
Department of Anatomy & Neurobiology, Reeve-Irvine Research Center, Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA, USA.
出版信息
J Neurosci Methods. 2010 Jun 30;190(1):63-70. doi: 10.1016/j.jneumeth.2010.04.025. Epub 2010 May 4.
PURPOSE
To develop three-dimensional (3D) constructs of retinal pigment epithelium (RPE) and early retina progenitor cells from human embryonic stem cells (hESCs).
METHODS
3D tissue constructs were developed by culturing hESC-derived neural retinal progenitors in a matrix on top of hESC-derived RPE cells in a cell culture insert. An osmolarity gradient maintained the nutrition of the 3D cell constructs. Cross-sections through hESC-derived tissue constructs were characterized by immunohistochemistry for various transcription factors and cell markers.
RESULTS
hESC-derived tissue constructs expressed transcription factors characteristic of retinal development, such as pax6, Otx2, Chx10, retinal RAX; Brn3b (necessary for differentiation of retinal ganglion cells); and crx and nrl (role in photoreceptor development). Many cells expressed neuronal markers including nestin, beta-tubulin and microtubule-associated proteins.
CONCLUSIONS
This study shows for the first time that 3D early retinal progenitor tissue constructs can be derived from hESCs.
目的
从人胚胎干细胞（hESC）中开发视网膜色素上皮（RPE）和早期视网膜祖细胞的三维（3D）构建体。
方法
通过在细胞培养插入物中，在 hESC 衍生的 RPE 细胞上的基质中培养 hESC 衍生的神经视网膜祖细胞，来开发 3D 组织构建体。渗透压梯度维持 3D 细胞构建体的营养。通过免疫组织化学对 hESC 衍生的组织构建体的横截面进行各种转录因子和细胞标志物的特征描述。
结果
hESC 衍生的组织构建体表达了视网膜发育特征的转录因子，如 pax6、Otx2、Chx10、视网膜 RAX；Brn3b（视网膜神经节细胞分化所必需的）；以及 crx 和 nrl（在光感受器发育中起作用）。许多细胞表达神经元标志物，包括巢蛋白、β-微管蛋白和微管相关蛋白。
结论
本研究首次表明可以从 hESC 中衍生出 3D 早期视网膜祖细胞组织构建体。
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