1 Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California , Davis, California.
J Ocul Pharmacol Ther. 2014 Mar-Apr;30(2-3):254-66. doi: 10.1089/jop.2013.0175. Epub 2014 Jan 23.
To support the growing promise of regenerative medicine in glaucoma, we characterized the similarities and differences between human trabecular meshwork (HTM) cells and human mesenchymal stem cells (hMSCs).
HTM cells and hMSCs were phenotypically characterized by flow cytometry. Using quantitative polymerase chain reaction, the expression of myoc, angptl7, sox2, pou5f1, and notch1 was determined in both cell types with and without dexamethasone (Dex). Immunosuppressive behavior of HTM cells and hMSCs was determined using T cells activated with phytohemagglutinin. T-cell proliferation was determined using BrdU incorporation and flow cytometry. Multipotency of HTM cells and hMSCs was determined using adipogenic and osteogenic differentiation media as well as aqueous humor (AH). Alpha-smooth muscle actin (αSMA) expression was determined in HTM cells, hMSCs, and HTM tissue.
Phenotypically, HTM and hMSCs expressed CD73, CD90, CD105, and CD146 but not CD31, CD34, and CD45 and similar sox2, pou5f1, and notch1 expression. Both cell types suppressed T-cell proliferation. However, HTM cells, but not hMSCs, upregulated myoc and angptl7 in response to Dex. Additionally, HTM cells did not differentiate into adipocytes or osteocytes. Culture of hMSCs in 20%, but not 100%, AH potently induced alkaline phosphatase activity. HTM cells in culture possessed uniformly strong expression of αSMA, which contrasted with the limited expression in hMSCs and spatially discrete expression in HTM tissue.
HTM cells possess a number of important similarities with hMSCs but lack multipotency, one of the defining characteristics of stem cells. Further work is needed to explore the molecular mechanisms and functional implications underlying the phenotypic similarities.
为了支持再生医学在青光眼领域的不断发展，我们对人眼小梁网（HTM）细胞和人间充质干细胞（hMSC）之间的相似性和差异进行了研究。
通过流式细胞术对 HTM 细胞和 hMSC 进行表型特征分析。通过定量聚合酶链反应（qPCR），检测了在加入和未加入地塞米松（Dex）的情况下，这两种细胞类型中 myoc、angptl7、sox2、pou5f1 和 notch1 的表达情况。使用植物血球凝集素激活的 T 细胞检测 HTM 细胞和 hMSC 的免疫抑制行为。通过 BrdU 掺入和流式细胞术检测 T 细胞的增殖情况。通过向脂肪和成骨分化培养基以及房水中添加诱导剂来检测 HTM 细胞和 hMSC 的多能性。检测 HTM 细胞、hMSC 和 HTM 组织中的α-平滑肌肌动蛋白（αSMA）表达情况。
表型上，HTM 和 hMSC 表达 CD73、CD90、CD105 和 CD146，但不表达 CD31、CD34 和 CD45，并且 Sox2、Pou5f1 和 Notch1 的表达也相似。这两种细胞类型均能抑制 T 细胞的增殖。然而，与 hMSC 不同的是，HTM 细胞在 Dex 作用下会上调 myoc 和 angptl7 的表达。此外，HTM 细胞不能分化为脂肪细胞或成骨细胞。将 hMSC 培养在 20%而不是 100%的房水中，可显著诱导碱性磷酸酶活性。在培养过程中，HTM 细胞均表现出强烈的 αSMA 表达，而 hMSC 则表达较弱，并且 HTM 组织中 αSMA 的表达呈空间离散分布。
HTM 细胞与 hMSC 具有许多重要的相似性，但缺乏多能性，这是干细胞的一个重要特征。需要进一步研究以探索这些表型相似性背后的分子机制和功能意义。
