Scaffold-Free Fabrication of Osteoinductive Cellular Constructs Using Mouse Gingiva-Derived Induced Pluripotent Stem Cells.
Scaffold-Free Fabrication of Osteoinductive Cellular Constructs Using Mouse Gingiva-Derived Induced Pluripotent Stem Cells.
复制标题
使用小鼠牙龈衍生的多能干细胞对骨诱导的细胞构建体的无脚手架构造。
DOI：
10.1155/2016/6240794
复制
发表时间：
2016
影响因子：
4.3
通讯作者：
Egusa H
中科院分区：
文献类型：
--
作者：
Okawa H;Kayashima H;Sasaki J;Miura J;Kamano Y;Kosaka Y;Imazato S;Yatani H;Matsumoto T;Egusa H
三维（3D）细胞构建体有望提供骨诱导材料，以开发用于骨再生的基于细胞的疗法。因此，诱导多能干细胞（iPSC）的增殖和自发聚集能力促使我们制造无支架iPSC构建体作为移植载体。将小鼠牙龈成纤维细胞衍生的iPSC（GF-iPSC）的胚状体接种在具有由温度响应性水凝胶制成的圆底孔的细胞室中。将收集的球状细胞构建体在成骨诱导培养基中培养30天，同时轻轻摇动，导致成骨标志物基因的显著上调。该结构由非结构化细胞团的内部区域和被成骨祖细胞样细胞包围的外部骨组织区域组成。外部骨组织被元素钙和磷以及羟基磷灰石强烈钙化。将GF-iPSC构建体皮下移植到免疫缺陷小鼠中有助于由畸胎瘤组织包围的广泛异位骨形成。这些结果表明，小鼠GF-iPSC可以促进骨诱导无支架3D细胞构建体的制造，其中钙化区域和周围的成骨细胞可以分别充当骨诱导的支架和驱动器。结合抑制畸胎瘤形成的技术，该系统可以为骨再生治疗提供有前途的策略。
Three-dimensional (3D) cell constructs are expected to provide osteoinductive materials to develop cell-based therapies for bone regeneration. The proliferation and spontaneous aggregation capability of induced pluripotent stem cells (iPSCs) thus prompted us to fabricate a scaffold-free iPSC construct as a transplantation vehicle. Embryoid bodies of mouse gingival fibroblast-derived iPSCs (GF-iPSCs) were seeded in a cell chamber with a round-bottom well made of a thermoresponsive hydrogel. Collected ball-like cell constructs were cultured in osteogenic induction medium for 30 days with gentle shaking, resulting in significant upregulation of osteogenic marker genes. The constructs consisted of an inner region of unstructured cell mass and an outer osseous tissue region that was surrounded by osteoblast progenitor-like cells. The outer osseous tissue was robustly calcified with elemental calcium and phosphorous as well as hydroxyapatite. Subcutaneous transplantation of the GF-iPSC constructs into immunodeficient mice contributed to extensive ectopic bone formation surrounded by teratoma tissue. These results suggest that mouse GF-iPSCs could facilitate the fabrication of osteoinductive scaffold-free 3D cell constructs, in which the calcified regions and surrounding osteoblasts may function as scaffolds and drivers of osteoinduction, respectively. With incorporation of technologies to inhibit teratoma formation, this system could provide a promising strategy for bone regenerative therapies.