PDF(Pubmed)
Abstract：
UNASSIGNED: Glass coverslips are used as a substrate since Harrison\'s initial nerve cell culture experiments in 1910. In 1974, the first study of brain cells seeded onto polylysine (PL) coated substrate was published. Usually, neurons adhere quickly to PL coating. However, maintaining cortical neurons in culture on PL coating for a prolonged time is challenging.
UNASSIGNED: A collaborative study between chemical engineers and neurobiologists was conducted to find a simple method to enhance neuronal maturation on poly-D-lysine (PDL). In this work, a simple protocol to coat PDL efficiently on coverslips is presented, characterized, and compared to a conventional adsorption method. We studied the adhesion and maturation of primary cortical neurons with various morphological and functional approaches, including phase contrast microscopy, immunocytochemistry, scanning electron microscopy, patch clamp recordings, and calcium imaging.
UNASSIGNED: We observed that several parameters of neuronal maturation are influenced by the substrate: neurons develop more dense and extended networks and synaptic activity is enhanced, when seeded on covalently bound PDL compared to adsorbed PDL.
UNASSIGNED: Hence, we established reproducible and optimal conditions enhancing maturation of primary cortical neurons in vitro. Our method allows higher reliability and yield of results and could also be profitable for laboratories using PL with other cell types.
UNASSIGNED: A collaborative study between chemical engineers and neurobiologists was conducted to find a simple method to enhance neuronal maturation on poly-D-lysine (PDL). In this work, a simple protocol to coat PDL efficiently on coverslips is presented, characterized, and compared to a conventional adsorption method. We studied the adhesion and maturation of primary cortical neurons with various morphological and functional approaches, including phase contrast microscopy, immunocytochemistry, scanning electron microscopy, patch clamp recordings, and calcium imaging.
UNASSIGNED: We observed that several parameters of neuronal maturation are influenced by the substrate: neurons develop more dense and extended networks and synaptic activity is enhanced, when seeded on covalently bound PDL compared to adsorbed PDL.
UNASSIGNED: Hence, we established reproducible and optimal conditions enhancing maturation of primary cortical neurons in vitro. Our method allows higher reliability and yield of results and could also be profitable for laboratories using PL with other cell types.
摘要：
■自1910年Harrison进行的初始神经细胞培养实验以来，玻璃盖玻片被用作底物。1974年，首次发表了将脑细胞接种到聚赖氨酸（PL）涂层基质上的研究。通常,神经元迅速粘附在PL涂层上。然而,在PL涂层上长时间维持培养中的皮质神经元是具有挑战性的。
■进行了化学工程师和神经生物学家之间的合作研究，以找到一种简单的方法来增强聚-D-赖氨酸（PDL）上的神经元成熟。在这项工作中,提出了一种在盖玻片上有效涂覆PDL的简单协议，characterized,并与传统的吸附方法进行了比较。我们用各种形态和功能方法研究了初级皮层神经元的粘附和成熟，包括相差显微镜，免疫细胞化学,扫描电子显微镜，膜片钳记录,和钙成像。
■我们观察到神经元成熟的几个参数受底物的影响：神经元发展出更密集和延伸的网络，突触活动增强，当在共价结合的PDL上接种时，与吸附的PDL相比。
■因此，我们建立了可重复和最佳的条件，以增强体外原代皮层神经元的成熟。我们的方法允许更高的可靠性和结果产量，并且对于使用PL和其他细胞类型的实验室也可能是有利可图的。
■进行了化学工程师和神经生物学家之间的合作研究，以找到一种简单的方法来增强聚-D-赖氨酸（PDL）上的神经元成熟。在这项工作中,提出了一种在盖玻片上有效涂覆PDL的简单协议，characterized,并与传统的吸附方法进行了比较。我们用各种形态和功能方法研究了初级皮层神经元的粘附和成熟，包括相差显微镜，免疫细胞化学,扫描电子显微镜，膜片钳记录,和钙成像。
■我们观察到神经元成熟的几个参数受底物的影响：神经元发展出更密集和延伸的网络，突触活动增强，当在共价结合的PDL上接种时，与吸附的PDL相比。
■因此，我们建立了可重复和最佳的条件，以增强体外原代皮层神经元的成熟。我们的方法允许更高的可靠性和结果产量，并且对于使用PL和其他细胞类型的实验室也可能是有利可图的。
