Multipotent Stem Cells from Trabecular Meshwork Become Phagocytic TM Cells
Multipotent Stem Cells from Trabecular Meshwork Become Phagocytic TM Cells
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DOI：
10.1167/iovs.11-9134
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发表时间：
2012-03-01
影响因子：
4.4
通讯作者：
Schuman, Joel S.
中科院分区：
文献类型：
--
作者：
Du, Yiqin;Roh, Danny S.;Schuman, Joel S.
目的。从人小梁网（TM）中分离和鉴定干细胞，并研究这些干细胞向TM细胞分化的潜力。人小梁网干细胞（TMSCs）通过荧光活化细胞分选或克隆培养分离为侧群细胞。通过免疫染色和定量RT-PCR将传代TMSCs与原代TM细胞进行比较。流式细胞术检测TMSC纯度，通过诱导神经细胞、脂肪细胞、角质细胞或TM细胞检测TMSC多能性。采用定量RT-PCR、免疫染色和免疫印迹检测差异基因表达。采用灭活的金黄色葡萄球菌生物颗粒，通过吞噬实验评估TM细胞的功能。侧群体和克隆分离细胞表达干细胞标记物ABCG2、Notch1、OCT-3/4、AnkG和MUC1，但不表达TM标记物AQP1、MGP、CHI3L1或TIMP3。传代的TMSCs是一个均匀的群体，95%的细胞对CD73、CD90、CD166或Bmi1呈阳性。通过表达神经标记物神经丝、β -微管蛋白III、GFAP， TMSCs表现出多能分化为多种细胞类型的能力；或角化细胞特异性标记物硫酸角蛋白和角化蛋白；或者脂肪细胞标志物ap2和瘦素。TMSC很容易分化为具有吞噬功能的TM细胞，表达了TM标记物AQP1、CHI3L1和timp3。作为侧群或克隆分离的TMSCs表达特定的干细胞标记物，具有同质性和多能性，能够分化为吞噬性TM细胞。这些细胞为青光眼的新型干细胞治疗提供了发展潜力。（Invest Ophthalmol Vis Sci. 2012;53:1566-1575） DOI:10.1167/iovs.11-9134
PURPOSE. To isolate and characterize stem cells from human trabecular meshwork (TM) and to investigate the potential of these stem cells to differentiate into TM cells.METHODS. Human trabecular meshwork stem cells (TMSCs) were isolated as side population cells by fluorescence-activated cell sorting or isolated by clonal cultures. Passaged TMSCs were compared with primary TM cells by immunostaining and quantitative RT-PCR. TMSC purity was assessed by flow cytometry and TMSC multipotency was examined by induction of neural cells, adipocytes, keratocytes, or TM cells. Differential gene expression was detected by quantitative RT-PCR, immunostaining, and immunoblotting. TM cell function was evaluated by phagocytic assay using inactivated Staphylococcus aureus bioparticles.RESULTS. Side population and clonal isolated cells expressed stem cell markers ABCG2, Notch1, OCT-3/4, AnkG, and MUC1 but not TM markers AQP1, MGP, CHI3L1, or TIMP3. Passaged TMSCs are a homogeneous population with > 95% cells positive to CD73, CD90, CD166, or Bmi1. TMSCs exhibited multipotent ability of differentiation into a variety of cell types with expression of neural markers neurofilament, beta-tubulin III, GFAP; or keratocyte-specific markers keratan sulfate and keratocan; or adipocyte markers ap2 and leptin. TMSC readily differentiated into TM cells with phagocytic function and expression of TM markers AQP1, CHI3L1, and TIMP3.CONCLUSIONS. TMSCs, isolated as side population or as clones, express specific stem cell markers, are homogeneous and multipotent, with the ability to differentiate into phagocytic TM cells. These cells offer a potential for development of a novel stem cell-based therapy for glaucoma. (Invest Ophthalmol Vis Sci. 2012;53:1566-1575) DOI:10.1167/iovs.11-9134