Production of a Human Growth Plate Organ-Chip Model of Skeletal Development

人体骨骼发育生长板器官芯片模型的制作

• 批准号: NC/X001873/1
• 负责人: Martin Knight
• 金额: $ 25.64万
• 依托单位: Queen Mary University of London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FX001873%2F1
• 关键词: Production; Human; Growth; Plate; Organ

The growth plate is an area of tissue at the ends of bones in children which determines the development of the skeleton. It represents a transition between bone and cartilage and is supported by blood vessels. Due to the importance of the growth plate, there is considerable interest in studying this organ and its role in development, ageing, and disease. To do so necessitates robust, reproducible and physiologically relevant experimental 'model' systems. Currently many studies rely on animals, typically mice and rats. Since mechanical forces are known to influence skeletal development in the body, some of the animal models include techniques to control mechanical forces on the growth plate whilst the animal is still alive. These widely used animal models often have limited reproducibility and fail to represent key behaviour in humans as well as prompting ethical concerns. There is therefore an urgent need to replace some of these animal models with non-animal, human experimental models. This would improve scientific rigour and relevance to human physiology as well as reducing the use of animals in science. Previously reported non-animal models of the growth plate fail to replicate the key features of the organ including a gradient of bone-cartilage tissue, mechanical loading and incorporation of blood vessels linking to the bone. Therefore, we will develop and validate a new human growth plate model using organ-on-a-chip technology. An organ-chip is an experimental model system in which human cells can be grown within interconnected channels through which fluid can be pumped providing the cells with the necessary nutrients to keep them alive. We will develop a growth plate organ-chip with the following key features: - A composite tissue graded from bone to cartilage - A channel replicating the blood vessels - Mechanical loading of the developing bone-cartilage growth plate- A reproducible, scalable system that can easily be adopted by other researchers To achieve the above, we will use human adult stem cells expanded from bone marrow. These stem cells will be grown within a 3D gel material in one of the channels in an organ-chip. The cells will then be differentiated into bone and cartilage cells using natural growth factors to create the graded bone-cartilage growth plate tissue. We will also create a blood vessel channel lined with human endothelial cells that form blood vessel walls. This blood vessel channel will provide a conduit for nutrient and hormone delivery to the developing growth plate as occurs in the body. To ensure reproducibility and scalability, the organ-chip model will be created with commercially available human stem cell and endothelial cells and within the commercially available organ-chip provided by Emulate Inc. This chip consists of two channels separated by a semi-permeable membrane allowing interaction between the developing bone-cartilage growth plate tissue in one channel and the blood vessel channel. In addition, we will utilise the ability of the Emulate organ-chip to provide controlled mechanical loading to the channels replicating the mechanical environment within the body. In this way, our vision is to create a highly reproducible and validated human vascularised growth plate organ-chip model which can be readily adopted by the scientific community reducing reliance on animal models and improving the quality of research into skeletal development in health and disease.

生长板是儿童骨骼末端的组织区域，这决定了骨骼的发展。它代表骨骼和软骨之间的过渡，并由血管支撑。由于生长板的重要性，研究器官及其在发育，衰老和疾病中的作用引起了很大的兴趣。这样做需要强大的，可重复的和生理上相关的实验“模型”系统。目前，许多研究依靠动物，通常是小鼠和大鼠。由于已知机械力会影响人体的骨骼发育，因此某些动物模型包括控制生长板上的机械力的技术，而动物仍然活着。这些广泛使用的动物模型通常具有有限的可重复性，并且无法代表人类的关键行为以及引起道德问题。因此，迫切需要用非动物的人类实验模型替代这些动物模型中的一些。这将改善科学严格和与人类生理学的相关性，并减少动物在科学中的使用。先前报道的生长板的非动物模型无法复制器官的关键特征，包括骨 - 骨折组织的梯度，机械负荷以及与骨骼相关的血管的掺入。因此，我们将使用Organ-A-A-Chip技术开发并验证新的人类生长板模型。器官芯片是一个实验模型系统，可以在该模型中生长人类细胞在相互联系的通道中生长，可以通过该模型泵送流体，从而为细胞提供必要的养分，以使其保持生存。我们将开发一个生长板器官芯片，具有以下关键特征： - 一种从骨骼到软骨分级的复合组织 - 复制血管的通道 - 正在发展的骨 - 骨折生长板的机械负载 - 一种可再现的可伸缩系统，可以轻松地由其他研究人员采用以上，以实现上述，我们将使用人类的成人干细胞从骨髓中扩展。这些干细胞将在器官芯片中的一个通道中的3D凝胶材料中生长。然后，使用自然生长因子将细胞分化为骨和软骨细胞，以创建分级的骨骼生长板组织。我们还将创建一个衬有人体内皮细胞的血管通道，形成血管壁。该血管通道将为体内发生的养分和激素递送到发育中的生长板提供管道。为了确保可重复性和可伸缩性，将使用市售的人类干细胞和内皮细胞以及在由仿真Inc提供的市售的器官芯片中创建器官芯片模型。该芯片由两个半渗透膜组成，该通道由一个可渗透的膜相互作用，可在一个通道和血液容器中允许发育中的骨骼生长板组织之间的相互作用。此外，我们将利用模拟器芯片的能力为复制人体机械环境的通道提供控制的机械载荷。这样，我们的愿景就是创建一个高度可再现和有效的人血管化生长板器官芯片模型，科学界很容易地采用降低对动物模型的依赖并提高健康和疾病骨骼发展的研究质量。