Subcutaneous Maturation of Neural Stem Cell-Loaded Hydrogels Forms Region-Specific Neuroepithelium
Subcutaneous Maturation of Neural Stem Cell-Loaded Hydrogels Forms Region-Specific Neuroepithelium
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DOI：
10.3390/cells7100173
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发表时间：
2018-10-01
期刊：
影响因子：
6
通讯作者：
Leipzig, Nic D.
中科院分区：
文献类型：
--
作者：
Farrag, Mahmoud;Leipzig, Nic D.
为了再生丢失的神经组织并最终恢复神经功能，可能需要一种整合干细胞并能够利用现有线索的组合方法。作为未来脊髓损伤治疗的第一步，本研究探讨了成体来源的神经干细胞(ANSCs)在生物材料构建物中的皮下成熟对aNSC分化和最终区域神经元特性的影响。为了实现这一点，我们将大鼠aNSCs包裹在壳聚糖管道内的壳聚糖基水凝胶中，该水凝胶带有固定的叠氮标记干扰素-Y。然后，我们将这些结构植入大鼠背部颈部、胸部和腰部的皮下组织，持续4、6和8周。收获支架后，用免疫组织化学方法和RT-qPCR方法对细胞分化进行定性和定量分析。结果显示，水凝胶支持aNSC在皮下环境中存活和分化长达4周，其标志是几种神经发生标志物的表达。最有趣的是，aNSCs表达与其植入区域相对应的区域特异性HOX基因。这项研究为进一步的翻译工作奠定了基础，以概括皮下组织中潜在的未被发现的模式线索，并为我们的生物工程方法可以形成尾状区域特异性神经上皮这一概念性前提提供支持。
A combinatorial approach integrating stem cells and capable of exploiting available cues is likely needed to regenerate lost neural tissues and ultimately restore neurologic functions. This study investigates the effects of the subcutaneous maturation of adult-derived neural stem cell (aNSCs) seeded into biomaterial constructs on aNSC differentiation and ultimate regional neuronal identity as a first step toward a future spinal cord injury treatment. To achieve this, we encapsulated rat aNSCs in chitosan-based hydrogels functionalized with immobilized azide-tagged interferon--y inside a chitosan conduit. Then, we implanted these constructs in the subcutaneous tissues in the backs of rats in the cervical, thoracic, and lumbar regions for 4, 6, and 8 weeks. After harvesting the scaffolds, we analyzed cell differentiation qualitatively using immunohistochemical analysis and quantitatively using RT-qPCR. Results revealed that the hydrogels supported aNSC survival and differentiation up to 4 weeks in the subcutaneous environment as marked by the expression of several neurogenesis markers. Most interesting, the aNSCs expressed region-specific Hox genes corresponding to their region of implantation. This study lays the groundwork for further translational work to recapitulate the potentially undiscovered patterning cues in the subcutaneous tissue and provide support for the conceptual premise that our bioengineering approach can form caudalized region-specific neuroepithelium.