Institute of Medicinal Plant Development , Chinese Academy of Medical Sciences and Peking Union Medical College , 151 Malianwa North Road , Haidian District, Beijing 100193 , China.
Department of Biochemistry and Molecular Medicine, School of Medicine , University of California , 2700 Stockton Blvd , Davis , California 95817 , United States.
ACS Appl Mater Interfaces. 2019 Feb 13;11(6):5821-5833. doi: 10.1021/acsami.8b19438. Epub 2019 Jan 29.
In this study, we established a long-term three-dimensional (3D) culture system by using integrin ligand modified alginate hydrogels to encapsulate and differentiate neural progenitor cells (NPCs) toward oligodendrocyte (OL) lineage cells. The porosity of the hydrogel was optimized by varying the alginate concentrations and then characterized by scanning electronic microscopy (SEM). The surface plasmon resonance (SPR) test was used to confirm the ligand-integrin interactions indicating adherence between the NPC surfaces and the hydrogels. Following encapsulation in the hydrogels, both mouse and human NPC sphere cultures could be maintained up to 90 days. Mouse NPC spheres were differentiated into viable neurons, astrocytes and mature OLs by day 60 in all groups whereas human NPC spheres were differentiated into neurons and later into GFAP positive astrocytes and O4 positive pre-OL within 90 days. The species difference in the timeline of OL development between mouse and human was reflected in this system. The ligand LXY30 interacting with the α3β1 integrin receptor was more effective in promoting the differentiation of hNPCs to OL lineage cells compared with the ligand LXW64 interacting with the αvβ3 integrin receptor, hyaluronic acid interacting with CD44 receptor or without any ligand. This study is the first to differentiate O4 pre-OLs from hNPCs in a LXY30-α3β1 (integrin-ligand) modified alginate 3D hydrogel culture. This 3D platform could serve as a valuable tool in disease modeling, drug discovery, and NPC transplantation.
在这项研究中，我们建立了一个长期的三维（3D）培养系统，使用整合素配体修饰的藻酸盐水凝胶来包封和分化神经祖细胞（NPC）向少突胶质细胞（OL）谱系细胞。通过改变藻酸盐浓度优化水凝胶的孔隙率，然后通过扫描电子显微镜（SEM）进行特征描述。表面等离子体共振（SPR）试验用于确认配体-整合素相互作用，表明 NPC 表面与水凝胶之间的粘附。在水凝胶包封后，鼠和人 NPC 球体培养物都可以维持长达 90 天。在所有组中，鼠 NPC 球体在第 60 天分化为有活力的神经元、星形胶质细胞和成熟的 OL，而人 NPC 球体在 90 天内分化为神经元，随后分化为 GFAP 阳性星形胶质细胞和 O4 阳性前 OL。这种系统反映了鼠和人之间 OL 发育的物种差异。与与 αvβ3 整合素受体相互作用的配体 LXW64、与 CD44 受体相互作用的透明质酸或没有任何配体相比，与 α3β1 整合素受体相互作用的配体 LXY30 更有效地促进 hNPC 向 OL 谱系细胞分化。这项研究首次在 LXY30-α3β1（整合素配体）修饰的藻酸盐 3D 水凝胶培养物中从 hNPC 分化出 O4 前 OL。这个 3D 平台可以作为疾病建模、药物发现和 NPC 移植的有价值的工具。
