Wnt5a-mediating neurogenesis of human adipose tissue-derived stem cells in a 3D microfluidic cell culture system
Wnt5a-mediating neurogenesis of human adipose tissue-derived stem cells in a 3D microfluidic cell culture system
复制标题
DOI：
10.1016/j.biomaterials.2011.05.090
复制
发表时间：
2011-10-01
期刊：
影响因子：
14
通讯作者：
Kang, Sookyung
中科院分区：
文献类型：
--
作者：
Choi, Jeein;Kim, Sohyeun;Kang, Sookyung
在干细胞生物学中，细胞可塑性是指干细胞分化成各种细胞谱系的能力。最近，细胞可塑性已被用于指给定细胞类型响应于特定刺激而可逆地去分化、再分化或转分化的能力。这些过程受到多种细胞内和细胞外生长和分化因子的调节，包括低氧。我们最近的研究表明，3D微流控细胞培养诱导hATSC（人脂肪组织来源的干细胞）中Wnt 5A/β-连环蛋白信号通路的激活。这导致hATSC自我更新和转分化为神经元。为了提高hATSCs在低氧和其他未知物理因素下的神经原性潜能，我们开发了一种无凝胶3D微流控细胞培养系统（3D-mu FCCS）。开发的功能结构的固定化的3D多细胞聚集体在微流体通道中，而不使用芯片上的矩阵。在芯片上生长的hATSC神经球的生长高于在培养皿中生长的对照细胞的生长。与从微流体通道层的中心主动迁移到微流体通道层的外部之后的对照细胞相比，在芯片系统中诱导分化导致神经元样细胞结构的诱导和TuJ或NF 160阳性长神经炎的呈现显著增加。我们还观察到，芯片神经发生系统诱导的GABA分泌神经元的水平显着较高，此外，几乎60%的细胞是GABA +细胞。最后，我们观察到，将每种细胞类型移植到小鼠SCI病变中1个月后，芯片培养和神经元分化的hATSC表现出以高比例有效转分化为NF 160+运动神经元的能力。有趣的是，我们的CHIP/PCR分析揭示了HIF 1 α诱导的芯片上的hATSC神经发生。这种诱导是HIF 1 α与细胞核中Oct 4和β-连环蛋白基因的调节区直接结合的结果。在我们开发的hATSCs的芯片培养中，诱导了低氧微环境。低氧水平诱导HIF 1 α表达，这通过HIF 1 α与β-连环蛋白和Oct 4的调节区的直接结合导致Wnt 5A/β-连环蛋白和Oct 4的表达增加。皇冠版权所有（C）2011由爱思唯尔有限公司出版。保留所有权利。
In stem cell biology, cell plasticity refers to the ability of stem cells to differentiate into a variety of cell lineages. Recently, cell plasticity has been used to refer to the ability of a given cell type to reversibly dedifferentiate, re-differentiate, or transdifferentiate in response to specific stimuli. These processes are regulated by multiple intracellular and extracellular growth and differentiation factors, including low oxygen. Our recent study showed that 3D microfluidic cell culture induces activation of the Wnt5A/beta-catenin signaling pathway in hATSCs (human Adipose Tissue-derived Stem Cells). This resulted in self renewal and transdifferentiation of hATSCs into neurons. To improve neurogenic potency of hATSCs in response to low oxygen and other unknown physical factors, we developed a gel-free 3D microfluidic cell culture system (3D-mu FCCS). The functional structure was developed for the immobilization of 3D multicellular aggregates in a microfluidic channel without the use of a matrix on the chip. Growth of hATSCs neurosphere grown on a chip was higher than the growth of control cells grown in a culture dish. Induction of differentiation in the Chip system resulted in a significant increase in the induction of neuronal-like cell structures and the presentation of TuJ or NF160 positive long neuritis compared to control cells after active migration from the center of the microfluidic channel layer to the outside of the microfluidic channel layer. We also observed that the chip neurogenesis system induced a significantly higher level of GABA secreting neurons and, in addition, almost 60% of cells were GABA + cells. Finally, we observed that 1 month of after the transplantation of each cell type in a mouse SCI lesion, chip cultured and neuronal differentiated hATSCs exhibited the ability to effectively transdifferentiate into NF160 + motor neurons at a high ratio. Interestingly, our CHIP/PCR analysis revealed that HIF1 alpha-induced hATSCs neurogenesis on the chip. This induction was a result of the direct binding of HIF1 alpha to the regulatory regions of the Oct4 and beta-catenin genes in nucleus. In the Chip culture of hATSCs that we developed, a low oxygen microenvironment was induced. The low oxygen level induced HIF1 alpha expression, which resulted in increased expression of Wnt5A/beta-catenin and Oct4 via the direct binding of HIF1 alpha to the regulatory regions of beta-catenin and Oct4. Crown Copyright (C) 2011 Published by Elsevier Ltd. All rights reserved.