Collaborative Research: RECODE: Microfluidic and genetic technologies to direct and select retinal cell types from human induced pluripotent stem cell-derived retinal organoids

细胞的分化是一系列复杂过程的结果。大多数干细胞分化方法会导致想要的和不想要的细胞类型的混合。更好地了解分化的关键分子控制点可以减少不良细胞的产生。这反过来可以提高基于细胞的治疗的有效性。基因和机械技术将被用来更好地理解导致视网膜细胞分化的途径。人们希望，这将导致退行性眼病的新疗法。生物制造业劳动力发展也将以几种方式得到支持。研究生将接受干细胞生物学和生物工程方面的培训，本科生将获得研究机会，并将通过细胞制造技术中心完成对行业的推广。控制人类诱导多能干细胞(HiPSC)的分化是一个挑战。细胞分子通路被编程为对外部信号做出反应。大多数协议在很大程度上依赖于自发的细胞命运承诺。这通常导致组织特定细胞类型的异质混合，其中许多对于再生细胞替代策略是不需要的，或者是有害的。应用微流控细胞分离和基因机械技术将评估它们可靠地将HiPSCs定向分化为视网膜细胞的能力。第一个目标是开发全基因组的方法，利用细胞分化和生物力学特性作为筛选，在视网膜细胞分化过程中针对所需的终点。这一目标可能会发现新的靶向途径，可以在培养过程中利用这些途径来调节视网膜的发生。第二个目标是利用系统生物学来识别视网膜细胞分化的主要调控因子，这些调控因子将通过观察这些通路对分化的视网膜亚型的调节作用而得到验证。第三个目标是使用细胞分离方法和新的标记物来提高棒和锥的适当比例的生产。这个RECODE项目由化学、生物工程、环境和运输系统部门的工程生物学和健康集群资助。这个奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The differentiation of cells is the result of a complicated set of processes. Most stem cell differentiation methods result in a mixture of desired and undesired cell types. Better understanding of key molecular control points of differentiation could decrease the generation of undesirable cells. That in turn could increase the effectiveness of cell-based therapies. Genetic and mechanical techniques will be employed to better understand the differentiation pathway resulting in retinal cells. The hope is that this will lead to new treatments for degenerative eye diseases. Biomanufacturing workforce development will also be supported in several ways. Graduate students will be trained in stem cell biology and bioengineering, undergraduate students will be provided research opportunities, and outreach to industry will be accomplished through the Center for Cell Manufacturing Technologies.Controlling human induced pluripotent stem cell (hiPSC) differentiation is a challenge. Cell molecular pathways are programmed to react to external cues. Most protocols depend to a great degree on spontaneous cell fate commitment. This often results in a heterogeneous mixture of tissue specific cell types, many of which are not needed, or detrimental, for regenerative cell replacement strategies. Applying microfluidic cell separation and geno-mechanical techniques will be evaluated for their ability to reliably direct differentiation of hiPSCs to retinal cells. The first objective is to develop genome-wide methods that use both cell differentiation and biomechanical properties as screens to target desired endpoints during retinal cell differentiation. This objective could uncover new target pathways that can be exploited during culture to regulate retinogenesis. The second objective is to use systems biology to identify master regulators of retinal cell differentiation These regulators will be validated by observing the effects of modulation of these pathways on differentiated retinal subtypes. A third objective is to use cell separation approaches coupled with novel markers to enhance the production of the proper proportion of rods to cones.This RECODE project is funded by the Engineering Biology and Health Cluster in the Division of Chemical, Bioengineering, Environmental, and Transport Systems.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.