Department of Anatomy and Neurobiology, Medical College of Virginia Campus, Virginia Commonwealth University, Richmond, VA, 23298, USA.
Department of Chemical and Life Science Engineering, College of Engineering, Virginia Commonwealth University, Richmond, Virginia, 23284, USA; Key Laboratory of Spine and Spinal Cord Injury Repair and Regeneration of Ministry of Education, Tongji Hospital, School of Medicine, Tongji University, Shanghai, 200065, PR China; School of Pharmaceutical Sciences, Guangzhou Medical University, Guangzhou, 511436, PR China.
J Neurosci Methods. 2020 Jun 1;339:108729. doi: 10.1016/j.jneumeth.2020.108729. Epub 2020 Apr 17.
Integrins are the major cell adhesion receptors expressed in almost all cell types connecting the extracellular matrix with cell cytoskeletons and transducing bi-directional signals across cell membranes. In the central nervous system (CNS), integrins are pivotal for CNS cell migration, differentiation, neurite outgrowth and synaptogenesis in both physiological and pathological conditions. Here we studied the effect of different integrin biding peptides for growth and development of primary cortical neurons in vitro.
Rat primary cortical neurons were cultured in an integrin-binding array platform, which contains immobilized varying short synthetic peptides that bind to 16 individual types of integrin on a 48-well cell culture plate. After cultured for 7 days, cells were fixed and processed for immunostaining with neuronal markers. The overall neuronal growth and neurite outgrowths were quantified.
We found that binding peptides for integrin αβ, αβ and αβ particularly the former two provided superior condition for neuronal growth, survival and maturation. Moreover, optimal neurite outgrowth was observed when neurons were cultured in 3-dimension using injectable hydrogel along with binding peptide for αβ or αβ integrins.
For primary neuronal culture, poly-D-lysine coating is conventional method to support cell attachment. Our study has demonstrated that selected integrin binding peptides provide greater support for the growth of cultured primary neurons.
These data suggest that integrin αβ and αβ are conducive for survival, growth and maturation of primary cortical neurons. This information could be utilized in designing combinational biomaterial and cell-based therapy for neural regeneration following brain injury.
整合素是几乎所有细胞类型中表达的主要细胞粘附受体，将细胞外基质与细胞细胞骨架连接起来，并在细胞膜两侧传递双向信号。在中枢神经系统（CNS）中，整合素对于 CNS 细胞在生理和病理条件下的迁移、分化、突起生长和突触发生至关重要。在这里，我们研究了不同整合素结合肽对体外原代皮质神经元生长和发育的影响。
将大鼠原代皮质神经元培养在整合素结合阵列平台上，该平台包含固定在 48 孔细胞培养板上的 16 种不同整合素结合的短合成肽。培养 7 天后，固定细胞并用神经元标志物进行免疫染色。定量分析总的神经元生长和突起生长。
我们发现，整合素 αβ、αβ 和 αβ 结合肽，特别是前两种，为神经元的生长、存活和成熟提供了更好的条件。此外，当神经元在可注射水凝胶中与 αβ 或 αβ 整合素结合肽一起培养时，观察到最佳的突起生长。
对于原代神经元培养，多聚-D-赖氨酸包被是支持细胞附着的常规方法。我们的研究表明，选择的整合素结合肽为培养的原代神经元的生长提供了更大的支持。
这些数据表明，整合素 αβ 和 αβ 有利于原代皮质神经元的存活、生长和成熟。这些信息可用于设计组合生物材料和基于细胞的治疗方法，以促进脑损伤后的神经再生。
