Identification of multipotent stem/progenitor cells in murine sclera.
Identification of multipotent stem/progenitor cells in murine sclera.
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DOI：
10.1167/iovs.11-7676
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发表时间：
2011-07
影响因子：
4.4
通讯作者：
Chia-Ling Tsai;Pei-Chang Wu;M. Fini;S. Shi
中科院分区：
文献类型：
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作者：
Chia-Ling Tsai;Pei-Chang Wu;M. Fini;S. Shi
目的巩膜形成眼球的纤维性外膜，并起支撑框架的作用。本研究的目的是检查巩膜是否含有间充质干/祖细胞。方法从C57 BL 6/J小鼠的视网膜和脉络膜分离巩膜组织，随后酶消化以释放单个细胞。通过BrdU标记、流式细胞术、逆转录-聚合酶链反应、免疫细胞化学和体内移植分析增殖能力、自我更新能力和多能分化能力。结果巩膜干/祖细胞（SSPCs）具有克隆形成能力和较高的增殖能力。这些细胞对间充质标志物Sca-1、CD90.2、CD 44、CD 105和CD 73呈阳性，对造血标志物CD 45、CD 11b、Flk 1、CD 34和CD 117呈阴性。除了表达干细胞基因ABCG 2、Six 2、Notch 1和Pax 6外，SSPC还能够分化为脂肪形成、软骨形成和神经形成谱系。结论巩膜中存在多能间充质干细胞。对SSPCs的深入研究有助于阐明巩膜炎、近视等巩膜疾病的细胞和分子机制。
PURPOSE The sclera forms the fibrous outer coat of the eyeball and acts as a supportive framework. The purpose of this study was to examine whether the sclera contains mesenchymal stem/progenitor cells. METHOD Scleral tissue from C57BL6/J mice was separated from the retina and choroid and subsequently enzyme digested to release single cells. Proliferation capacity, self-renewal capacity, and ability for multipotent differentiation were analyzed by BrdU labeling, flow cytometry, reverse transcriptase-polymerase chain reaction, immunocytochemistry, and in vivo transplantation. RESULTS The scleral stem/progenitor cells (SSPCs) possessed clonogenic and high doubling capacities. These cells were positive for the mesenchymal markers Sca-1, CD90.2, CD44, CD105, and CD73 and negative for the hematopoietic markers CD45, CD11b, Flk1, CD34, and CD117. In addition to expressing stem cell genes ABCG2, Six2, Notch1, and Pax6, SSPCs were able to differentiate to adipogenic, chondrogenic, and neurogenic lineages. CONCLUSIONS This study indicates that the sclera contains multipotent mesenchymal stem cells. Further study of SSPCs may help elucidate the cellular and molecular mechanism of scleral diseases such as scleritis and myopia.