Cerebrospinal fluid replacement solutions promote neuroglia migratory behaviors and spinal explant outgrowth in microfluidic culture
Cerebrospinal fluid replacement solutions promote neuroglia migratory behaviors and spinal explant outgrowth in microfluidic culture
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DOI：
10.1002/term.3164
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发表时间：
2020-12-15
影响因子：
3.3
通讯作者：
Vazquez, Maribel
中科院分区：
文献类型：
--
作者：
Cliver, Richard N.;Ayers, Brian;Vazquez, Maribel
神经系统疾病是创伤性损伤、遗传疾病或慢性健康状况的后果，影响着全世界数百万成年人。当代研究已经开始将神经胶质细胞纳入新兴的神经神经系统疗法中，以利用脑和脊髓中神经胶质介导的突触的再生潜力。然而，脑脊液（CSF）包围神经胶质细胞并与其相关突触界面的作用仍然只是部分探索。脑脊液在蛛网膜下腔（SAS）内的流动循环必需的多肽、代谢物和生长因子，通过健康胶质的信号传导直接影响神经反应和恢复。尽管人工脑脊液溶液可用于神经外科和NS治疗，但组织工程项目仍在继续使用细胞培养基，如Neurobasal （NB）和Dulbecco的Modified Eagle Medium (DMEM)，用于开发和表征许多可移植细胞、基质和集成细胞系统。目前的研究检测了神经胶质雪旺细胞（ShC）和脊髓外植体（SCE）在脑脊液替代溶液（Elliott's B solution， EBS）中的体外行为，该溶液广泛用于治疗神经系统疾病。我们的测试使用EBS在胶质系（gLL）微流控装置内创建细胞外因子的定义化学微环境，之前由我们的团队描述。gLL在规模上与体内的SAS相当，后者包裹内源性CSF并通过对流扩散机制实现分子运输。我们的研究结果表明，EBS溶液促进了ShC的存活、形态和增殖，与传统DMEM中测量的结果相似，此外还支持脑源性生长因子（BDNF）对胶质趋化行为的响应。我们的数据表明，ShC对BDNF的高浓度和低浓度梯度具有显著的趋化性，在EBS和DMEM溶液稀释剂中形成的梯度具有统计学差异。此外，用EBS溶液培养的SCE有助于测量与报道的体内测量相适应的神经突外植体延伸。这些数据强调了结合CSF替代液来询问细胞行为和推进再生NS治疗的翻译意义和优势。
Disorders of the nervous system (NS) impact millions of adults, worldwide, as a consequence of traumatic injury, genetic illness, or chronic health conditions. Contemporary studies have begun to incorporate neuroglia into emerging NS therapies to harness the regenerative potential of glial-mediated synapses in the brain and spinal cord. However, the role of cerebrospinal fluid (CSF) that surrounds neuroglia and interfaces with their associated synapses remains only partially explored. The flow of CSF within subarachnoid spaces (SAS) circulates essential polypeptides, metabolites, and growth factors that directly impact neural response and recovery via signaling with healthy glia. Despite the availability of artificial CSF solutions used in neurosurgery and NS treatments, tissue engineering projects continue to use cell culture media, such as Neurobasal (NB) and Dulbecco's Modified Eagle Medium (DMEM), for development and characterization of many transplantable cells, matrixes, and integrated cellular systems. The current study examined in vitro behaviors of glial Schwann cells (ShC) and spinal cord explants (SCE) within a CSF replacement solution, Elliott's B Solution (EBS), used widely in the treatment of NS disorders. Our tests used EBS to create defined chemical microenvironments of extracellular factors within a glial line (gLL) microfluidic device, previously described by our group. The gLL is comparable in scale to the in vivo SAS that envelopes endogenous CSF and enables molecular transport via mechanisms of convective diffusion. Our results illustrate that EBS solutions facilitate ShC survival, morphology, and proliferation similar to those measured in traditional DMEM, and additionally support glial chemotactic behaviors in response to brain-derived growth factor (BDNF). Our data indicates that ShC undergo significant chemotaxis toward high and low concentration gradients of BDNF with statistical differences between gradients formed within diluents of EBS and DMEM solutions. Moreover, SCE cultured with EBS solutions facilitated measurement of neurite explant extension commensurate with reported in vivo measurements. This data highlights the translational significance and advantages of incorporating CSF replacement fluids to interrogate cellular behaviors and advance regenerative NS therapies.