Cox2-mediated PGE2 production via p38/JNK-c-fos signaling inhibits cell apoptosis in 3D floating culture clumps of mesenchymal stem cell/extracellular matrix complexes
Cox2-mediated PGE2 production via p38/JNK-c-fos signaling inhibits cell apoptosis in 3D floating culture clumps of mesenchymal stem cell/extracellular matrix complexes
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DOI：
10.1016/j.bbrc.2020.05.100
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发表时间：
2020-09-17
影响因子：
3.1
通讯作者：
Kurihara, Hidemi
中科院分区：
文献类型：
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作者：
Komatsu, Nao;Kajiya, Mikihito;Kurihara, Hidemi
间充质干细胞（Mesenchymal stem cells，MSCs）是一类成体干细胞，由于其自我更新特性、多能性和营养因子产生而引起科学和医学的关注。虽然最初在经典的二维（2D）塑料板上研究MSC，但广泛的科学努力已经开发出三维（3D）MSC培养系统，包括可以模拟物理条件的MSC球状体和类器官。此外，我们最近开发了用于新型骨再生细胞疗法的MSC/细胞外基质（ECM）复合物（C-MSC）的3D培养团块。值得注意的是，尽管广泛接受细胞从培养板脱离导致细胞凋亡，所谓的失巢凋亡，但这些3D MSC构建体可以维持在漂浮培养条件下。目前，尚不清楚为什么3D漂浮培养的MSC构建体可以逃避失巢凋亡。为了回答这个问题，本研究探讨了三维漂浮培养的C-MSCs中营养因子的产生，这些营养因子在体外对失巢凋亡起细胞保护作用，并阐明了潜在的分子机制。与二维培养板相比，三维漂浮培养C-MSCs中COX 2介导的PGE 2的产生显著增加，COX 2的抑制可显著诱导细胞凋亡。在C-MSC的制备过程中，细胞片层从培养板上脱落激活了p38/JNK-c-Fos信号通路。此外，通过化学抑制剂阻断该信号传导废除了COX 2/PGE 2表达并诱导严重的细胞凋亡。这些结果表明，细胞脱离通过p38/JNK-c-Fos信号转导促进细胞保护性COX 2介导的PGE 2合成，揭示了允许漂浮培养的3D MSC构建体抵抗失巢凋亡的可能机制。(C)2020爱思唯尔公司All rights reserved.
Mesenchymal stem cells (MSCs), a class of adult stem cells, have attracted scientific and medical attention due to their self-renewing properties, multipotency, and trophic factor production. Although MSCs were originally studied on classical two-dimensional (2D) plastic plates, extensive scientific efforts have developed three-dimensional (3D) MSC culture systems, including MSCs spheroids and organoids that can mimic physical conditions. Moreover, we have recently developed 3D culture clumps of MSCs/extracellular matrix (ECM) complexes (C-MSCs) for novel bone regenerative cell therapy. Of note, even though it is widely accepted that cell detachment from the culture plate causes cell apoptosis, so called anoikis, these 3D MSCs constructs can be maintained in floating culture conditions. Currently, it is unclear why 3D floating-cultured MSCs constructs can escape from anoikis. To answer this question, the present study explored trophic factor production in 3D floating-cultured C-MSCs that play a cytoprotective role against anoikis and clarified the underlying molecular mechanism in vitro. Compared with cells cultured on 2D plastic plates, PGE2 production mediated by COX2 was significantly increased, and its inhibition drastically induced cell apoptosis in 3D floating-cultured C-MSCs. In the process of C-MSCs preparation, detachment of the cell sheet from culture plate activated the p38/JNK-c-Fos signaling pathway. Moreover, blockage of this signaling by chemical inhibitors abrogated COX2/PGE2 expressions and induced severe apoptosis. These results demonstrated that cell detachment facilitates cytoprotective COX2-mediated PGE2 synthesis via p38/JNK-c-Fos signaling, revealing a possible mechanism that allows resistance against anoikis in floating-cultured 3D MSCs constructs. (C) 2020 Elsevier Inc. All rights reserved.