Quantitative Evaluation of Human Umbilical Vein and Induced Pluripotent Stem Cell-Derived Endothelial Cells as an Alternative Cell Source to Skin-Specific Endothelial Cells in Engineered Skin Grafts.
Quantitative Evaluation of Human Umbilical Vein and Induced Pluripotent Stem Cell-Derived Endothelial Cells as an Alternative Cell Source to Skin-Specific Endothelial Cells in Engineered Skin Grafts.
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对人脐静脉和诱导多能干细胞衍生的内皮细胞作为工程皮肤移植物中皮肤特异性内皮细胞的替代细胞来源的定量评估。
DOI：
10.1089/wound.2020.1163
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发表时间：
2021
影响因子：
4.9
通讯作者：
Abaci,HasanErbil
中科院分区：
文献类型：
--
作者：
Pappalardo,Alberto;Herron,Lauren;AlvarezCespedes,DavidE;Abaci,HasanErbil
目的：比较人脐静脉内皮细胞(HUVECs)、诱导多能干细胞(IPSC)来源的内皮细胞(IECs)和人真皮血液内皮细胞(HDBECs)在体外和免疫缺陷小鼠体内有效地血管化工程化人皮肤结构(HSCs)的能力。我们用实时定量聚合酶链式反应(RT-qPCR)检测2D培养和HSCs中关键的内皮、真皮和表皮基因的表达。通过苏木精-伊红染色和IF染色评价表皮的完整性和增殖。结果：IF证实了IEC对内皮表型的承诺。RT-qPCR显示HUVECs和IECs与HDBEC相比未成熟；体外，HDBECs和HUVECs的血管网络伸展相似，尽管血管直径不同，但IECs形成了无组织的基本管状结构；在体内，所有ECs都产生了不同尺寸的离散血管网络。HUVECs和HDBECs维持较高的基底层角质形成细胞增殖。HDBECs对细胞外基质的表达以及真皮的增殖和分化的影响最大。创新：本研究首次对HDBECs、HUVECs和IECs在HSC中的血管生成性能进行了直接和定量的比较。结论：HUVECs和IECs可以作为HDBEC的替代细胞来源，以促进无血运工程移植物的短期存活。然而，HDBEC保持其毛细血管特性，并在促进真皮和表皮成熟方面优于其他类型的EC。HDBECs的这些固有特性可能会影响皮肤移植物的长期功能。
Objective:We compared the capability of human umbilical vein endothelial cells (HUVECs), induced pluripotent stem cell (iPSC)-derived endothelial cells (iECs), and human dermal blood endothelial cells (HDBECs) to effectively vascularize engineered human skin constructs (HSCs)in vitroand on immunodeficient mice.Approach:We quantified the angiogenesis within HSCs bothin vitroandin vivothrough computational analyses of immunofluorescent (IF) staining. We assayed with real-time quantitative PCR (RT-qPCR) the expression of key endothelial, dermal, and epidermal genes in 2D culture and HSCs. Epidermal integrity and proliferation were also evaluated through haematoxylin and eosin staining, and IF staining.Results:IF confirmed iEC commitment to endothelial phenotype. RT-qPCR showed HUVECs and iECs immaturity compared with HDBECs.In vitro, the vascular network extension was comparable for HDBECs and HUVECs despite differences in vascular diameter, whereas iECs formed unorganized rudimentary tubular structures.In vivo, all ECs produced discrete vascular networks of varying dimensions. HUVECs and HDBECs maintained a higher proliferation of basal keratinocytes. HDBECs had the best impact on extracellular matrix expression, and epidermal proliferation and differentiation.Innovation:To our knowledge, this study represents the first direct and quantitative comparison of HDBECs, HUVECs, and iECs angiogenic performance in HSCs.Conclusions:Our data indicate that HUVECs and iECs can be an alternative cell source to HDBEC to promote the short-term viability of prevascularized engineered grafts. Nevertheless, HDBECs maintain their capillary identity and outperform other EC types in promoting the maturation of the dermis and epidermis. These intrinsic characteristics of HDBECs may influence the long-term function of skin grafts.