Engineering microscale hydrogel deposition to direct single stem cell differentiation

工程微型水凝胶沉积指导单干细胞分化

• 批准号: 10582026
• 负责人: Jae-Won Shin
• 金额: $ 25万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10582026
• 关键词: 3-Dimensional; Address; Biocompatible Materials; Biophysics; Bone Regeneration; Cell Therapy; Cell Volumes; Cells; Clinical; Clinical Trials; Cues; Data; Deposition; Disease; Elasticity; Encapsulated; Engineering; Formulation; Gel; Genetic; Genetic Transcription; Growth; Heterogeneity; Hydrogels; Individual; Integrins; Ion Channel; Ligands; Mechanics; Membrane; Mesenchymal Stem Cells; Natural regeneration; Outcome; Population; Property; Reproducibility; Testing; Therapeutic; Thinness; Tissues; adult stem cell; base; clinical efficacy; clinically relevant; design; improved; in vivo; injured; insight; multidisciplinary; osteogenic; programs; public health relevance; regenerative; stem cell differentiation; stem cell fate; stem cell function; stem cell growth; stem cells; therapy outcome; tissue regeneration

PROJECT SUMMARY Adult stem cells hold broad-ranging clinical potential to regenerate injured tissues. For instance, mesenchymal stem cells (MSCs) have been investigated in over 950 clinical trials for use in many disease indications. Despite their significant clinical relevance, however, there is currently lack of the mechanistic understanding to precisely control MSC functions for reproducible therapeutic outcomes. Engineered hydrogels have been used to reveal the ability of MSCs to sense and respond to matrix biophysical cues, which subsequently impact the differentiation potential of MSCs. However, leveraging these insights for therapeutic purposes has been challenging, since current approaches to interface a cell population with a hydrogel by uncontrolled mixing overlook the significance of heterogeneity in the local amount of the gel presented to individual cells, leading to variable and unclear cell-material interactions at the single cell level. We describe herein a highly efficient approach to control microscale hydrogel deposition around single cells in a 3D space independently of gel composition and elasticity. Using this approach, our preliminary data show that MSCs rapidly expand in volume when they adhere to an integrin ligand in thinner gels. We show that encapsulating single MSCs in a thin gel coating is sufficient to enhance the osteogenic potential of MSCs even when gel elasticity is low. We will build upon these results to test the hypothesis that controlling local gel deposition around single MSCs impacts membrane tension and lineage specification by regulating cell volume expansion. In Aim 1, we will determine the effect of varying local gel deposition on regulatory volume decrease by modulating mechanosensitive ion channels and its impact on membrane tension of MSCs. In Aim 2, we will determine how varied local gel deposition impacts single MSC fate and MSC-based bone regeneration. We predict that there exists a transcriptional program that is selectively activated when the gel deposition becomes thinner, thereby impacting lineage specification of MSCs independently of gel elasticity. The project is highly multidisciplinary in that it will employ a combination of expertise in biomaterials, biophysical, genetic, and in vivo approaches to address the specific aims. The results will help to define local gel deposition as an important determinant of stem cell growth, thereby impacting stem cell mechanics and fate. Given the clinical relevance of these cells, our results will inform formulation design of MSC-based therapeutics for improved regenerative outcomes.

项目概要 成体干细胞具有广泛的临床潜力，可以再生受损组织。例如， 间充质干细胞 (MSC) 已在超过 950 项临床试验中进行了研究，用于 许多疾病的指征。然而，尽管它们具有重要的临床意义，但目前 缺乏精确控制 MSC 功能以实现可重复性的机制理解 治疗结果。工程水凝胶已被用来揭示 MSC 的能力 感知并响应基质生物物理线索，从而影响分化 间充质干细胞的潜力。然而，利用这些见解来达到治疗目的已经 具有挑战性，因为目前将细胞群与水凝胶连接的方法是通过 不受控制的混合忽​​视了凝胶局部量的异质性的重要性 呈现给单个细胞，导致细胞与材料之间的相互作用可变且不明确 单细胞水平。我们在此描述了一种控制微型水凝胶的高效方法 3D 空间中单细胞周围的沉积，与凝胶成分和弹性无关。 使用这种方法，我们的初步数据表明，当间充质干细胞进入细胞时，其体积会迅速扩大。 在较薄的凝胶中粘附于整合素配体。我们证明将单个 MSC 封装在薄层中 即使凝胶弹性较低，凝胶涂层也足以增强 MSC 的成骨潜力。 低的。我们将根据这些结果来检验控制局部凝胶沉积的假设 单个 MSC 周围通过调节细胞影响膜张力和谱系规范 体积膨胀。在目标 1 中，我们将确定不同的局部凝胶沉积对 通过调节机械敏感离子通道减少调节体积及其对 MSC 的膜张力。在目标 2 中，我们将确定不同的局部凝胶沉积如何影响 单一 MSC 命运和基于 MSC 的骨再生。我们预测存在一个 当凝胶沉积变薄时选择性激活的转录程序， 从而影响 MSC 的谱系规格，与凝胶弹性无关。该项目是 高度多学科性，因为它将结合生物材料方面的专业知识， 生物物理、遗传和体内方法来解决特定目标。结果会有所帮助 将局部凝胶沉积定义为干细胞生长的重要决定因素，从而影响 干细胞力学和命运。鉴于这些细胞的临床相关性，我们的结果将告知 基于 MSC 的疗法的配方设计，以改善再生结果。