Scaff-Net: 3 Dimensional multiphoton polymerisation printed scaffolds for medium throughput recording from stem cell derived human cortical networks.

神经和神经退行性疾病是对人类健康、生活质量和社会日益严重的主要威胁。欧盟每年治疗脑部疾病的费用估计约为7980亿欧元，抑郁症、双相情感障碍、癫痫和痴呆等脑部疾病影响了约1.65亿欧洲人，即三分之一的人（欧盟委员会）。有必要对这种情况进行药物治疗和治疗，然而，由于缺乏合适的方法来测试和筛选潜在有用的分子，进展受到阻碍。一个主要问题是，传统上大多数研究都是用动物组织进行的，比如大鼠或小鼠的大脑，或者从这些动物身上培养的神经元。然而，似乎人类脑细胞的反应及其在许多方面的联系可能与小鼠神经元不同。另一个问题是，虽然细胞可以在几周或几个月的时间内培养和研究，但它们通常是在平坦的表面上生长，以供科学家和制药行业进行测试，这与它们在大脑中的生长方式完全不同。人类的大脑是由神经元网络组成的，每个神经元通过连接相互交流，这些连接在三维空间中向四面八方辐射。连接模式及其复杂性是人类大脑运作的基本特征之一。在二维平面上培养的神经网络不能像在活体大脑中那样建立连接，因此它们产生的活动也会有所不同。为了能够开发针对人类疾病的药物和疗法，理想的细胞系统将是人类神经网络的三维培养，我们可以记录其活动并测试药物。如果可以同时从不同的网络进行许多记录，这样可以在更短的时间内测试许多药物，那将是理想的。这就是我们将通过脚手架网实现的目标。使用基于激光的技术，我们将“打印”支持神经网络的小支架。为了获得人类神经网络，我们将使用来自皮肤样本的人类干细胞。我们将从干细胞中培养神经元，并在支架上培养它们，在那里它们将形成连接的网络。如今，许多二维平面培养物生长在许多小电极的网格上，以记录活动。在scaffold - net中，我们将在这些多电极阵列网格的每个单个电极上打印单个支架和神经元网络。这意味着这些电极阵列将从能够在单个平面2D培养物上记录多个位点转变为能够同时记录支架上多个3D网络（约60个！）的活动。这将意味着药物测试可以更快地进行，因此治疗发现将更快。在未来，我们期望scaffold - net设备对药物发现的方式产生重大影响，并改变该领域。

A major growing threat to human health, quality of life and society is that of neurological and neurodegenerative diseases. The annual cost of treating brain conditions in the EU is estimated at ~798 billion Euros, with brain disorders such as depression, bipolar disorder, epilepsy and dementia affecting about 165 million Europeans or 1 in 3 people (European Commission).There is a need for drug treatments and therapies for such conditions, however, advance is hampered by the lack of suitable methods to test and screen potentially useful molecules. One major problem is that most research traditionally has been conducted using animal tissues such as rat or mouse brains or cultured neurons from these animals. However, it seems that human brain cells responses and their connections in many ways may be different to mouse neurons. Another problem is that although cells can be cultured and investigated over a period of weeks or months they are usually grown on flat surfaces for testing by scientists and the pharmaceutical industry, which is totally different to the way that they grow in the brain. The human brain consists of networks of neurons communicating with each through connections that radiate in all directions in 3 dimensions. The pattern of connections and its complexity is one of the fundamental features that enables human brains to function as they do. Cultures of neuronal networks grown on flat 2D surfaces cannot make the same connections as they would in the living brain, and therefore the activity that they produce will also be different. To be able to develop drugs and therapies for human conditions, the ideal cell system would be 3 dimensional cultures of human neuronal networks that we could record activity from and test drugs. It would also be ideal if many recordings could be done at the same time from different networks so that many drugs could be tested in a shorter time. This is what we will achieve with Scaff-Net. Using laser-based technology we will "print" small scaffolds that will support neuronal networks. To obtain human neuronal networks we will use human derived stem cells that come from skin samples. We will grow neurons from the stem cells and culture these on the scaffolds where they will form connected networks. Many of the 2D flat cultures grown today are grown on grids of many small electrodes to record activity. In Scaff-Net we will print a single scaffold and neuronal network on each single electrode of these grids of multi electrode arrays. This will mean that these electrode arrays will be transformed from being able to record at many sites on a single flat 2D culture to be able to record activity from many 3D networks on scaffolds (about 60!) at the same time. This will mean that the testing of drugs can be made quicker, and so therapeutic discoveries will be made quicker. In the future we would expect Scaff-Net devices to have a major impact on the way that drug discovery is conducted and transform the field.