Fibroblasts derived from human embryonic stem cells direct development and repair of 3D human skin equivalents.
Fibroblasts derived from human embryonic stem cells direct development and repair of 3D human skin equivalents.
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源自人类胚胎干细胞的成纤维细胞直接发育和修复 3D 人类皮肤等同物。
DOI：
10.1186/scrt51
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发表时间：
2011-02-21
影响因子：
7.5
通讯作者：
Garlick JA
中科院分区：
文献类型：
--
作者：
Shamis Y;Hewitt KJ;Carlson MW;Margvelashvilli M;Dong S;Kuo CK;Daheron L;Egles C;Garlick JA
多能的人类干细胞作为祖细胞和终末分化细胞的来源，在未来的再生治疗中有着巨大的应用前景。然而，这些治疗将依赖于新方法的发展，这些方法可以最好地评估多潜能干细胞衍生细胞的组织结果，并且对于更好地预测体内移植后的安全性和稳定性将是必不可少的。在这项研究中，我们使用工程化的人类皮肤等效物(HSE)作为平台来表征从人类胚胎干细胞(HES)中衍生出来的成纤维细胞。我们鉴定了两个不同的具有间充质细胞特性的HES来源的细胞系(EDK和H9-MSC)的表型和分泌谱，并比较了它们在诱导分化为骨和脂肪以及在它们被整合到工程化的HSE的间质中后的生物学潜力。虽然EDK和H9-MSC细胞株具有相似的形态和间充质细胞标志物的表达，但当它们被整合到HSE的间质中时，它们表现出不同的功能特性。EDK细胞显示出真皮成纤维细胞的特征，可以支持上皮组织的发育，并使使用皮肤伤口愈合的3D组织模型产生的伤口重新上皮化，这与肝细胞生长因子(HGF)的产生有关。慢病毒shRNA介导的HGF敲除导致EDK细胞HGF分泌显著减少，导致其促进角质形成细胞增殖和皮肤创面再上皮化的能力显着降低。相反，H9-MSCs表现出间充质干细胞(MSC)的特征，但不具有真皮成纤维细胞的特征，因为它们在单层培养中经历了多向分化，但无法支持上皮组织的发育和修复，并产生显著较低水平的HGF。我们的发现表明，HES来源的细胞可以定向到特定的和可替代的间充质细胞命运，其功能可以在工程化HSE中区分开来。HES来源的间充质细胞在3D工程化HSE中的表征证明了该组织平台在治疗移植之前预测HES来源的成纤维细胞的功能特性的实用性。
Pluripotent, human stem cells hold tremendous promise as a source of progenitor and terminally differentiated cells for application in future regenerative therapies. However, such therapies will be dependent upon the development of novel approaches that can best assess tissue outcomes of pluripotent stem cell-derived cells and will be essential to better predict their safety and stability following in vivo transplantation. In this study we used engineered, human skin equivalents (HSEs) as a platform to characterize fibroblasts that have been derived from human embryonic stem (hES) cell. We characterized the phenotype and the secretion profile of two distinct hES-derived cell lines with properties of mesenchymal cells (EDK and H9-MSC) and compared their biological potential upon induction of differentiation to bone and fat and following their incorporation into the stromal compartment of engineered, HSEs. While both EDK and H9-MSC cell lines exhibited similar morphology and mesenchymal cell marker expression, they demonstrated distinct functional properties when incorporated into the stromal compartment of HSEs. EDK cells displayed characteristics of dermal fibroblasts that could support epithelial tissue development and enable re-epithelialization of wounds generated using a 3D tissue model of cutaneous wound healing, which was linked to elevated production of hepatocyte growth factor (HGF). Lentiviral shRNA-mediated knockdown of HGF resulted in a dramatic decrease of HGF secretion from EDK cells that led to a marked reduction in their ability to promote keratinocyte proliferation and re-epithelialization of cutaneous wounds. In contrast, H9-MSCs demonstrated features of mesenchymal stem cells (MSC) but not those of dermal fibroblasts, as they underwent multilineage differentiation in monolayer culture, but were unable to support epithelial tissue development and repair and produced significantly lower levels of HGF. Our findings demonstrate that hES-derived cells could be directed to specified and alternative mesenchymal cell fates whose function could be distinguished in engineered HSEs. Characterization of hES-derived mesenchymal cells in 3D, engineered HSEs demonstrates the utility of this tissue platform to predict the functional properties of hES-derived fibroblasts before their therapeutic transplantation.
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影响因子：
15.8
作者：
Barberi T;Willis LM;Socci ND;Studer L
通讯作者：
Studer L
影响因子：
4
作者：
Jiang, Xiaohua;Gwye, Ynnez;Lawlor, Elizabeth R.
通讯作者：
Lawlor, Elizabeth R.
影响因子：
6.1
作者：
Chen, Hsin-Fu;Chuang, Ching-Yu;Kuo, Hung-Chih
通讯作者：
Kuo, Hung-Chih
影响因子：
6.5
作者：
GARLICK, JA;TAICHMAN, LB
通讯作者：
TAICHMAN, LB
DOI：
10.1073/pnas.0704421104
发表时间：
2007-06-26
影响因子：
11.1
作者：
Ortiz, Luis A.;DuTreil, Maria;Phinney, Donald G.
通讯作者：
Phinney, Donald G.