High-throughput stem cell bioprocess design - environmental influences on pluripotent stem cell expansion and differentiation

再生医学是一个快速发展的价值数十亿美元的多学科领域，是医学、工程和科学的交叉点。它的目标是创造活的、功能性的组织，可以修复或替换受损、患病或老化的组织或器官。干细胞对再生医学治疗的成功至关重要，我们需要的是弥合基础生物学和干细胞临床应用之间差距的技术。拟议的研究计划将加速这些技术的发展，特别是以一种可靠、可重复的方式培养干细胞来构建组织。研究的重点是多能干细胞，因为它们有可能在体内产生任何类型的细胞，即大量的特化（分化）细胞。它们可以来源于早期发育的囊胚（胚胎干细胞）的内部细胞群，也可以来源于通过添加关键因子修饰的皮肤细胞（诱导多能干细胞）等成体细胞。有效再生医学治疗道路上的主要障碍之一是可获得的供体细胞数量有限，以及培养和分化细胞的小规模、劳动密集型方法。该研究的主要目标是开发在计算机控制的生物反应器中有效培养人类多能干细胞的工程方法和技术。首先，关键的生物反应器条件（如氧气水平）和营养物质的运输的影响将检查个别干细胞群和整个生物反应器。其次，将开发新技术，通过使用多个较小的生物反应器作为高通量工具来快速研究许多条件，从而加速生物过程的发展。长期成果包括创新的生物工艺设计，以及关于输出参数（即细胞类型、行为）如何受到细胞加工参数（即生物反应器氧水平）影响的关键知识。这对于干细胞技术的监管批准和临床应用至关重要，包括用于治疗人类疾病（如中风、帕金森病、糖尿病）的细胞/组织的产生和移植。

Regenerative medicine is a rapidly growing billion dollar multidisciplinary field at the intersection of medicine, engineering and science. It aims to create living, functional tissues that can repair or replace damaged, diseased or aging tissues or organs. Stem cells are critical to the success of regenerative medicine therapies, and what are needed are technologies to bridge the gap between basic biology and the clinical application of stem cells. The proposed research program will accelerate the development of these technologies, specifically targeting a reliable, reproducible way to grow stem cells to build tissues. The research will focus on pluripotent stem cells as they have the potential to generate any cell type in the body, i.e. large numbers of specialized (differentiated) cells. They can be derived from the inner cell mass of an early development blastocyst (embryonic stem cells) or from adult cells such as skin cells that are modified by the addition of key factors to be pluripotent (induced pluripotent stem cells). One of the major stumbling blocks on the road to effective regenerative medicine treatments is the limited number of available donor cells, and the small-scale, labour-intensive methods for culturing and differentiating them. The primary objective of the proposed research is to develop engineering methods and technology for efficiently growing human pluripotent stem cells in computer-controlled bioreactors. First, the effect of key bioreactor conditions (such as oxygen level) and the transport of nutrients will be examined for individual stem cell clusters and entire bioreactors. Second, new technology will be developed to accelerate the development of bioprocesses through the use of multiple smaller bioreactors as high-throughput tools to study many conditions quickly. The long term outcomes include innovative bioprocess designs, as well as critical knowledge of how output parameters (i.e. cell type, behaviour) are influenced by cell processing parameters (i.e. bioreactor oxygen level). This is fundamentally necessary for the regulatory approval and clinical application of stem cell technologies, including the generation and transplantation of cells/tissues for the treatment of human ailments (i.e. stroke, Parkinson's disease, diabetes).