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

这个小型企业创新研究(SBIR)项目的更广泛的影响/商业潜力是为生物医学研究社区提供新的工具，用于下一代组织培养平台，包括那些来自患者样本的平台。如果成功，这项技术将使研究人员能够以一种与当前实验室基础设施广泛兼容的格式实现3D细胞培养技术的自动化和标准化。它将在商业生物制造/制药开发和学术最先进之间架起一座桥梁。此外，拟议的技术将为科学界提供新的工具，以扰乱和观察单个细胞如何相互作用及其与微环境的相互作用。在单细胞分辨率下探索生物学特征是当前生物医学研究的主要趋势，从癌症治疗、诊断和免疫学的临床应用，到全球基础科学倡议，如人类细胞图谱项目。这笔赠款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第一阶段项目的智力优势是开发一种用于单细胞分辨率的有机体或其他3D组织培养的自动化仪器。有机化合物有可能改变个性化的药物和疾病模型，但目前的培养方法控制不力，导致个体样本和实验室之间的差异很大。人们认识到，细胞接种参数的标准化、重复性和严格控制对于在临床上从有机物中提取预测值将是至关重要的，特别是当需要许多统计重复时。开发的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亚纳升液体体积内的单个细胞。目的是评估实施拟议的细胞打印技术的可行性，以完全控制每个有机细胞中组成细胞的类型和数量来播种有机细胞培养物。使用Lgr5+干细胞来源的肠道器官作为模型培养系统，将开发出从原始组织中制备“准备好打印”的输入单个干细胞的方案。然后，计划通过改变细胞数量、细胞类型和细胞外基质成分来评估细胞播种状态对有机体形态的影响，以检验通过高含量成像评估的受控播种将导致形态重复性改善的假设。该奖项反映了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 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.