Data_Sheet_1_Mechanosensitivity of Human Oligodendrocytes.PDF

Oligodendrocytes produce and repair myelin, which is critical for the integrity and function of the central nervous system (CNS). Oligodendrocyte and oligodendrocyte progenitor cell (OPC) biology is modulated in vitro by mechanical cues within the magnitudes observed in vivo. In some cases, these cues are sufficient to accelerate or inhibit terminal differentiation of murine oligodendrocyte progenitors. However, our understanding of oligodendrocyte lineage mechanobiology has been restricted primarily to animal models to date, due to the inaccessibility and challenges of human oligodendrocyte cell culture. Here, we probe the mechanosensitivity of human oligodendrocyte lineage cells derived from human induced pluripotent stem cells. We target phenotypically distinct stages of the human oligodendrocyte lineage and quantify the effect of substratum stiffness on cell migration and differentiation, within the range documented in vivo. We find that human oligodendrocyte lineage cells exhibit mechanosensitive migration and differentiation. Further, we identify two patterns of human donor line-dependent mechanosensitive differentiation. Our findings illustrate the variation among human oligodendrocyte responses, otherwise not captured by animal models, that are important for translational research. Moreover, these findings highlight the importance of studying glia under conditions that better approximate in vivo mechanical cues. Despite significant progress in human oligodendrocyte derivation methodology, the extended duration, low yield, and low selectivity of human-induced pluripotent stem cell-derived oligodendrocyte protocols significantly limit the scale-up and implementation of these cells and protocols for in vivo and in vitro applications. We propose that mechanical modulation, in combination with traditional soluble and insoluble factors, provides a key avenue to address these challenges in cell production and in vitro analysis.

少突胶质细胞（Oligodendrocytes）能够合成并修复髓磷脂，这对中枢神经系统（central nervous system, CNS）的完整性与功能至关重要。少突胶质细胞及其前体细胞（oligodendrocyte progenitor cell, OPC）的生物学特性在体外受到机械信号的调控，这类信号的强度范围与体内观测到的水平一致。在部分情况下，此类机械信号足以加速或抑制小鼠少突胶质前体细胞的终末分化。然而迄今为止，由于人类少突胶质细胞培养存在可及性差与技术挑战，我们对少突胶质细胞谱系力学生物学的认知主要局限于动物模型。本研究针对由人类诱导多能干细胞（human induced pluripotent stem cells）分化得到的人类少突胶质细胞谱系细胞的机械敏感性展开探究。我们靶向人类少突胶质细胞谱系中表型各异的不同发育阶段，并在体内已报道的基质刚度范围内，量化了基质刚度对细胞迁移与分化的影响。研究发现，人类少突胶质细胞谱系细胞表现出机械敏感性的迁移与分化行为。进一步地，我们鉴定出两种依赖人类供体细胞系的机械敏感性分化模式。我们的研究结果揭示了人类少突胶质细胞应答的个体差异——这类差异无法通过动物模型捕捉，且对转化研究具有重要价值。此外，这些研究结果强调了在更贴近体内机械信号的条件下研究神经胶质细胞（glia）的重要性。尽管人类少突胶质细胞的诱导分化方法已取得显著进展，但现有人类诱导多能干细胞来源少突胶质细胞的制备方案仍存在周期长、产量低、选择性差等缺陷，极大限制了这类细胞及相关方案在体内与体外研究中的规模化应用与推广。我们提出，将机械调控与传统的可溶性、非可溶性因子调控相结合，可为解决细胞生产与体外分析中的上述挑战提供关键可行路径。