Systematic stem cell bioprocess engineering

基于细胞制造的生物过程工程代表了生物治疗发展的下一个浪潮。这些产品是否能进入市场取决于以稳健和经济有效的方式制造电池的技术。干细胞作为制造细胞的“原材料”发挥着重要作用，然而，产品是活细胞这一事实提出了一系列独特的制造和控制挑战。干细胞必须被引导到合适的路径来产生所需的细胞类型。多能干细胞（PSC）是一个强大的系统，因为它们可以在培养中无限扩增，同时在适当的条件下保留分化成几乎任何成熟细胞类型的潜力。它们作为电池制造的“通用来源”提供了巨大的希望。为了实现这一承诺，生物工程基础包括生物反应器设计、过程控制和计算建模必须与发育生物学和3D组织组织的知识相结合。这些因素指导了我们这项研究计划的长期目标，即开发创新的基于细胞的技术，以促进PSC的自组织功能行为，用于制造血细胞产品。我们将专注于开发微加工，生物反应器和其他生物工程技术，以强有力地控制三维系统中细胞分化的时空组织。了解导致不对称细胞命运决定的空间和时间信号事件将使我们能够确定生物过程控制点，我们可以利用这些控制点来促进从PSC到体外分化细胞的理想分化轨迹。然而，这最终将使细胞生产规模扩大，通常涉及异质细胞群体，其中竞争的生长速度和培养要求产生不断变化和不稳定的培养，需要控制策略。我们将在PSC中设计合成生物学电路，以整合多个输入并驱动复杂的基因表达程序，以可扩展的方式指导血液诱导和分化过程中的细胞命运，以达到我们期望的终点。这代表了一种新的生物过程控制层，超越了可溶性因子添加的标准方法。实时、动态和剂量控制的信号因子将创造一个更均匀的培养微环境，从而提高我们指导分化的能力，确保靶细胞扩增，减少输出变异性。拟议的研究计划将开发技术，使新兴的加拿大电池制造业，并将支持培训一个多样化的bb60合格的研究人员群体，使用公平和包容性的招聘和指导实践。这些人员将非常适合为高价值的下一代工业生物技术制造工艺做出贡献。

Bioprocess engineering of cell based manufacturing represents a next wave in the development of biologic treatments. Whether or not these products ever reach the market depends upon technologies to manufacture cells in a robust and cost effective manner. Stem cells play an important role as the `raw material' in manufacturing cells, however the fact that the product is living cells brings forward a unique set of manufacturing and control challenges. Stem cells must be directed down the appropriate path to generate the cell types needed. Pluripotent stem cells (PSC) are a powerful system as they can be expanded in culture indefinitely while retaining the potential to differentiate into almost any mature cell type when placed under the right conditions. They offer immense promise as a `universal source' for cell manufacturing. To realize this promise, bioengineering fundamentals including bioreactor design, process control, and computational modelling must be combined with knowledge of developmental biology and 3D tissue organisation. These factors instruct our long-term goal of this research program which is to develop innovative cell based technologies to promote self-organized functional behaviour from PSC for manufacturing blood cell products. We will focus on developing microfabrication, bioreactor, and other bioengineering technologies to robustly control spatial temporal organization of cell differentiation in 3D systems. Understanding the spatial and temporal signaling events leading to asymmetric cell fate decisions will allow us to identify bioprocess control points that we can use to promote desirable differentiation trajectories to go from PSC to differentiated cells in vitro. This will ultimately enable scaled-up cell production, however, often that involves heterogeneous cell populations where competing growth rates and culture requirements yield continuously changing and unstable cultures, necessitating control strategies. We will engineer synthetic biology circuits in PSC to integrate multiple inputs and drive complex gene expression programs to direct cell fate during blood induction and differentiation, toward our desired end point in a scalable way. This represents a novel layer of bioprocess control, beyond standard approaches of soluble factor addition. Real-time, dynamic, and dose-control of signaling factors will create a more homogeneous culture microenvironment, thereby improving our ability to direct differentiation, ensure target cell expansion, and reduce output variability. The proposed research program will develop technologies that enable the emerging Canadian cell manufacturing industry, and will support the training of a diverse group of >6 qualified researchers, using equitable and inclusive recruitment and mentoring practices. These personnel will be ideally suited to contribute to high value next-generation industrial biotechnology manufacturing processes.