SBIR Phase I: Controlled Organoid Synthesis with Fluorescence-Activated Cell Printing

这项小型企业创新研究（SBIR）项目的更广泛影响/商业潜力是为生物医学研究界提供下一代组织培养平台的新工具，包括来自患者样本的组织培养平台。如果成功，这项技术将使研究人员能够以与当前实验室基础设施广泛兼容的格式自动化和标准化3D细胞培养技术。它将弥合商业生物制造/制药开发与学术前沿之间的差距。此外，提出的技术将为科学界提供新的工具来干扰和观察单个细胞如何相互作用以及与微环境的相互作用。从癌症治疗、诊断和免疫学的临床应用，到人类细胞图谱项目等全球基础科学倡议，以单细胞分辨率探测生物特征支撑着当前生物医学研究的主要趋势。这项资助的目标是促进单细胞处理仪器的商业化，用于研究型大学和制药研发实验室。这个SBIR一期项目的智力优势是开发一种自动化仪器，用于播种类器官或其他单细胞分辨率的3D组织培养。类器官具有改变个性化医疗和疾病建模的潜力，但目前的培养方法控制不善，导致个体样本和实验室之间的差异很大。人们认识到，标准化、可重复性和严格控制细胞播种参数对于在临床中从类器官中提取预测值至关重要，特别是当需要进行许多统计重复时。该技术是一种高分辨率的机器人细胞打印仪器，能够快速和确定地播种含有亚纳升液体体积的单个细胞。目标是评估实施所提出的细胞打印技术的可行性，以种子类器官培养，完全控制每个类器官中组成细胞的类型和数量。使用Lgr5+干细胞衍生的肠道类器官作为模型培养系统，将开发从原代组织中制备“可打印”输入的单个干细胞的方案。然后，我们的计划是通过改变细胞数量、细胞类型和ECM成分来评估细胞播种状态对类器官形态的影响，以验证通过高含量成像评估的受控播种将提高形态再现性的假设。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is to provide the biomedical research community with a new tool for next-generation tissue culture platforms, including those derived from patient samples. If successful, this technology will enable researchers to automate and standardize 3D cell culture techniques in a format that is broadly compatible with current laboratory infrastructure. It would bridge a gap between commercial biomanufacturing/pharmaceutical development and the academic state-of-the-art. Moreover, the proposed technology will provide the scientific community with novel tools to perturb and observe how individual cells interact with each other and with their microenvironment. Probing biological signatures at single-cell resolution underpins major current trends in biomedical research, from clinical applications in cancer therapy, diagnostics, and immunology, to global basic science initiatives such as the Human Cell Atlas project. The goal of this grant is to facilitate commercialization of a single-cell handling instrument for use in research universities and pharmaceutical R&D laboratories.The intellectual merit of this SBIR Phase I project is to develop an automated instrument for seeding organoids or other 3D tissue cultures with single-cell resolution. Organoids have the potential to transform personalized medicine and disease modelling, but current culture methods are poorly controlled, leading to wide variability between individual samples and across labs. It is recognized that standardization, reproducibility, and tight control of cell seeding parameters will be paramount for drawing predictive value from organoids in the clinic, particularly when many statistical replicates are required. The technology developed is a high-resolution robotic cell printing instrument capable of rapidly and deterministically seeding single cells contained within sub-nanoliter liquid volumes. The goal is to evaluate the feasibility of implementing the proposed cell printing technology to seed organoid cultures with complete control of the type and number of constituent cells in each organoid. Using Lgr5+ stem cell-derived intestinal organoids as a model culture system, protocols will be developed for preparing "printer-ready" input single stem cells from primary tissue. Then, the plan is to evaluate the effects of cell seeding state on organoid morphology by varying cell number, cell type and ECM components, to test the hypothesis that controlled seeding will result in improved morphological reproducibility, as assessed by high-content imaging.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.