Human endometrial stromal stem cells differentiate into megakaryocytes with the ability to produce functional platelets.
Human endometrial stromal stem cells differentiate into megakaryocytes with the ability to produce functional platelets.
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DOI：
10.1371/journal.pone.0044300
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Chen Y
中科院分区：
文献类型：
--
作者：
Wang J;Chen S;Zhang C;Stegeman S;Pfaff-Amesse T;Zhang Y;Zhang W;Amesse L;Chen Y
人子宫内膜是一种含有子宫内膜间质干细胞（endometrial stromal stem cells，hESSCs）的高动态组织。hESSC已经分化成许多细胞谱系。然而，尚未研究hESSC向巨核细胞（MK）的分化。本研究的目的是研究从hESSC产生MK和随后产生功能性血小板（PLT）的可行性。在我们的研究中，hESSC从子宫内膜基质细胞培养，通过阳性基质细胞特异性标志物（CD 90和CD 29）和阴性造血干细胞标志物（CD 45和CD 34）表达证实。然后，hESSC在补充有血小板生成素（TPO）的培养基中分化18天。流式细胞术和共聚焦显微镜分析MK分化。收集分化培养基，通过流式细胞术、透射电子显微镜和功能测量进行PLT产生分析。我们的结果显示：1）通过特异性标志物的表达鉴定，成功地从hESSC产生MK（CD 41 a：1±0.09%和39±3.0%; CD 42 b：1.2±0.06%和28± 2.0%，对照与分化），伴有多能转录因子减少（Oct 4和Sox 2）表达; 2）分化培养基中的PLT水平为16±1个/μl，由大小（2-4 μm）和CD 41 a表达确定（CD 41 a：1±0.4%和90± 2.0%，对照vs.分化）; 3）生成的PLT具有功能性，如凝血酶刺激后CD 62 p表达和纤维蛋白原结合的上调所证明; 4）通过电子显微镜分析测定，释放的PLT显示出与来自外周血的PLT相似的超微结构特征（α颗粒、空泡和致密管状系统）。数据证明了从hESSC产生MK的可行性，并且所产生的MK释放功能性PLT。因此，hESSCs可能是一个潜在的新的干细胞来源，在体外MK/PLT生产。
Human endometrium is a high dynamic tissue that contains endometrial stromal stem cells (hESSCs). The hESSCs have been differentiated into a number of cell lineages. However, differentiation of hESSCs into megakaryocytes (MKs) has not yet been investigated. The aim of this study was to investigate the feasibility of MK generation from hESSCs and subsequent production of functional platelets (PLTs). In our study, hESSCs were cultured from endometrial stromal cells as confirmed by positive stromal cell specific markers (CD90 and CD29) and negative hematopoietic stem cell markers (CD45 and CD34) expression. Then, hESSCs were differentiated in a medium supplemented with thrombopoietin (TPO) for 18 days. The MK differentiation was analyzed by flow cytometry and confocal microscopy. The differentiation medium was collected for PLT production analysis by flow cytometry, transmission electron microscopy and functional measurements. Our results show: 1) MKs were successfully generated from hESSCs as identified by expression of specific markers (CD41a: 1±0.09% and 39±3.0%; CD42b: 1.2±0.06% and 28±2.0%, control vs. differentiation) accompanied with reduction of pluripotent transcription factors (Oct4 and Sox2) expression; 2) The level of PLTs in the differentiation medium was 16±1 number/µl as determined by size (2–4 µm) and CD41a expression (CD41a: 1±0.4% and 90±2.0%, control vs. differentiation); 3) Generated PLTs were functional as evidenced by the up-regulation of CD62p expression and fibrinogen binding following thrombin stimulation; 4) Released PLTs showed similar ultra-structure characteristics (alpha granules, vacuoles and dense tubular system) as PLTs from peripheral blood determined by electron microscopic analysis. Data demonstrate the feasibility of generating MKs from hESSCs, and that the generated MKs release functional PLTs. Therefore, hESSCs could be a potential new stem cell source for in vitro MK/PLT production.
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影响因子：
5.3
作者：
Wolff EF;Gao XB;Yao KV;Andrews ZB;Du H;Elsworth JD;Taylor HS
通讯作者：
Taylor HS
影响因子：
3.6
作者：
Gargett, Caroline E.;Schwab, Kjiana E.;Wu, Di
通讯作者：
Wu, Di
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2.6
作者：
Shim, MH;Hoover, A;Reems, JA
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金属蛋白酶调节可改善小鼠胚胎干细胞的体外产生有效的血小板。
影响因子：
15.3
作者：
Nishikii, Hidekazu;Eto, Koji;Tamura, Noriko;Hattori, Koichi;Heissig, Beate;Kanaji, Taisuke;Sawaguchi, Akira;Goto, Shinya;Ware, Jerry;Nakauchi, Hiromitsu
通讯作者：
Nakauchi, Hiromitsu
影响因子：
4
作者：
Matthai, C;Horvat, R;Kolbus, A
通讯作者：
Kolbus, A