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.

项目总结 成体干细胞具有广泛的再生受损组织的临床潜力。例如, 间充质干细胞(MSCs)已经在950多项临床试验中进行了研究，用于 许多疾病的迹象。然而，尽管它们具有重要的临床相关性，但目前有 缺乏对精确控制MSC功能以实现可重复性的机械理解 治疗效果。工程水凝胶已被用来揭示骨髓间充质干细胞 感知和响应基质生物物理线索，这些线索随后会影响分化 间充质干细胞的潜能。然而，将这些见解用于治疗目的一直是 具有挑战性，因为目前将细胞群体与水凝胶相结合的方法是通过 不加控制的混合忽视了凝胶局部含量的不均一性的重要性 呈现给单个细胞，导致细胞-材料在 单细胞水平。我们在这里描述了一种控制微尺度水凝胶的高效方法。 在3D空间中的单个细胞周围沉积，与凝胶组成和弹性无关。 使用这种方法，我们的初步数据显示，当MSCs 在更薄的凝胶中粘着整合素配体。我们展示了将单个MSCs封装在一个薄的 凝胶涂层足以增强MSCs的成骨能力，即使在凝胶弹性 很低。我们将在这些结果的基础上测试控制局部凝胶沉积的假设 单个MSCs周围通过调节细胞影响膜张力和谱系特征 成交量扩大。在目标1中，我们将确定不同的局部凝胶沉积对 调节机械敏感离子通道的调节容量减少及其对心脏功能的影响 骨髓间充质干细胞的膜张力。在目标2中，我们将确定不同的局部凝胶沉积如何影响 单个MSC的命运和基于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， 从而独立于凝胶弹性影响MSCs的谱系特性。该项目是 高度多学科，因为它将结合生物材料方面的专业知识， 生物物理、遗传和活体方法来解决特定的目标。结果将会有所帮助 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。考虑到这些细胞的临床相关性，我们的结果将告诉我们 用于改善再生结果的基于MSC的疗法的配方设计。