Self-assembly of differentiated progenitor cells facilitates spheroid human skin organoid formation and planar skin regeneration.
Self-assembly of differentiated progenitor cells facilitates spheroid human skin organoid formation and planar skin regeneration.
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分化祖细胞的自组装促进了球体人皮肤手机体形成和平面皮肤再生。
DOI：
10.7150/thno.59661
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发表时间：
2021
期刊：
影响因子：
12.4
通讯作者：
Strunk D
中科院分区：
文献类型：
--
作者：
Ebner-Peking P;Krisch L;Wolf M;Hochmann S;Hoog A;Vári B;Muigg K;Poupardin R;Scharler C;Schmidhuber S;Russe E;Stachelscheid H;Schneeberger A;Schallmoser K;Strunk D
单细胞自组装实体器官将极大地扩展再生医学的适用性。干细胞/祖细胞可以自我组织成微型器官单位，称为类器官，部分模拟组织功能和再生。在这里，我们展示了在人血小板裂解液的支持下，成体和诱导多能干细胞（iPSC）衍生的成纤维细胞、角化细胞和内皮祖细胞在体内三维自组装成平面人类皮肤和体外一种新型球形皮肤类器官。方法：从人体组织中分离原代内皮集落形成细胞（ECFCs）、皮肤成纤维细胞（FBs）和角质形成细胞（KCs），并在二维无异种条件下进行多克隆繁殖。根据效率优化方案，将人组织来源的iPSCs分化为内皮细胞（hiPSC-ECs）、成纤维细胞（hiPSC-FBs）和角质形成细胞（hiPSC-KCs）。流式细胞术证实了细胞的身份和纯度，克隆原性显示了它们的干细胞/祖细胞潜力。含有培养条件的人血小板来源的生长因子促进了三细胞型浮动球体的形成，使用纳米颗粒细胞标记来监测组织过程。在免疫缺陷小鼠全层创面移植hipsc来源的单细胞悬浮液后，评估了平面人皮肤再生。结果：类器官显示出独特的结构，表面固定的角质形成细胞围绕着基质核心，并在炎症刺激下表现出特定的信号模式。需要FGF-7 mRNA转染才能加速角质形成细胞的长期适应性。在成人或ipsc衍生的皮肤细胞悬浮液移植后两周内，分层的人类皮肤也能自我组装，愈合小鼠的深度伤口。在共移植内皮祖细胞的存在下，移植血管化明显加快。从机制上讲，细胞外囊泡介导了血小板衍生的多因子营养效应。异种移植无肿瘤发生。结论：这说明了优越的祖细胞自组织原理，并为漂浮球形皮肤类器官提供了新的快速3D皮肤相关药物高含量测试机会。多细胞移植自组织促进了利用人血小板因子支持的细胞悬浮移植的基于ipsc的器官再生策略的发展。
Self-assembly of solid organs from single cells would greatly expand applicability of regenerative medicine. Stem/progenitor cells can self-organize into micro-sized organ units, termed organoids, partially modelling tissue function and regeneration. Here we demonstrated 3D self-assembly of adult and induced pluripotent stem cell (iPSC)-derived fibroblasts, keratinocytes and endothelial progenitors into both, planar human skin in vivo and a novel type of spheroid-shaped skin organoids in vitro, under the aegis of human platelet lysate. Methods: Primary endothelial colony forming cells (ECFCs), skin fibroblasts (FBs) and keratinocytes (KCs) were isolated from human tissues and polyclonally propagated under 2D xeno-free conditions. Human tissue-derived iPSCs were differentiated into endothelial cells (hiPSC-ECs), fibroblasts (hiPSC-FBs) and keratinocytes (hiPSC-KCs) according to efficiency-optimized protocols. Cell identity and purity were confirmed by flow cytometry and clonogenicity indicated their stem/progenitor potential. Triple cell type floating spheroids formation was promoted by human platelet-derived growth factors containing culture conditions, using nanoparticle cell labelling for monitoring the organization process. Planar human skin regeneration was assessed in full-thickness wounds of immune-deficient mice upon transplantation of hiPSC-derived single cell suspensions. Results: Organoids displayed a distinct architecture with surface-anchored keratinocytes surrounding a stromal core, and specific signaling patterns in response to inflammatory stimuli. FGF-7 mRNA transfection was required to accelerate keratinocyte long-term fitness. Stratified human skin also self-assembled within two weeks after either adult- or iPSC-derived skin cell-suspension liquid-transplantation, healing deep wounds of mice. Transplant vascularization significantly accelerated in the presence of co-transplanted endothelial progenitors. Mechanistically, extracellular vesicles mediated the multifactorial platelet-derived trophic effects. No tumorigenesis occurred upon xenografting. Conclusion: This illustrates the superordinate progenitor self-organization principle and permits novel rapid 3D skin-related pharmaceutical high-content testing opportunities with floating spheroid skin organoids. Multi-cell transplant self-organization facilitates development of iPSC-based organ regeneration strategies using cell suspension transplantation supported by human platelet factors.
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影响因子：
5.8
作者：
Gu, Zuguang;Eils, Roland;Schlesner, Matthias
通讯作者：
Schlesner, Matthias
影响因子：
17.1
作者：
Hu, Shiqi;Li, Zhenhua;Cheng, Ke
通讯作者：
Cheng, Ke
影响因子：
--
作者：
Borena, Bizunesh M.;Meyer, Evelyne;Spaas, Jan H.
通讯作者：
Spaas, Jan H.
影响因子：
5.9
作者：
Brok-Volchanskaya, Vera S.;Bennin, David A.;Slukvin, Igor
通讯作者：
Slukvin, Igor
影响因子：
4.6
作者：
Hochmann S;Mittermeir M;Santic R;Koszik F;Griessner L;Sonderegger AS;Hoffmann T;Russe E;Scheiblhofer S;Weiss R;Mandler M;Schneeberger A;Strunk D
通讯作者：
Strunk D