Chemically Defined and Biologically Active Microcarriers for Cell Expansion

用于细胞扩增的化学成分明确且具有生物活性的微载体

基本信息

批准号：
1709179
负责人：
Padma Gopalan
金额：
$ 39万
依托单位：
University of Wisconsin-Madison
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1709179&HistoricalAwards=false
关键词：
Chemically Defined Biologically Active Microcarriers

项目摘要

PART 1: NON-TECHNICALOver the past three decades, a variety of fundamental scientific discoveries have identified cells that can be used in medical research and therapy. However, in order to be effectively used, these cells must be manufactured efficiently, controllably, and reproducibly. Culturing and expanding cells while enhancing their desired function is essential for biomanufacturing, and critical for ultimate use of cells to understand and treat human disease. This proposal focuses on biomanufacturing two important cell types, namely human mesenchymal stem cells (hMSCs) that have been used in over 500 human clinical trials, and endothelial cells (ECs) that form human blood vessels. Developing a better in vitro understanding and control over regulation of hMSC and EC behaviors such as adhesion, proliferation, and differentiation may facilitate their efficient manufacturing and increased success as cell therapies. This research program will lead to understanding of cell behaviour, and to more effective manufacturing of therapeutic cells, by developing biomaterials to mimic parts of the native extracellular matrix (ECM). These biomaterials in the form of polymer coatings will aid in answering key fundamental questions in cell biology and provide an efficient, well-defined platform for cell biomanufacturing. The resulting polymer-coated "microcarriers" for 3D cell expansion can be delivered to cell biologists and bioengineers, who can then customize them to probe key biological questions. Hands-on exhibits developed on Stem Cells and Tissue Engineering through this research will be disseminated nationally by the PI's and the graduate students. The research program will serve to inspire and involve undergraduate students in polymer science and engineering research, by working with graduate student mentors. PART 2: TECHNICAL SUMMARYThis research program will study the synthesis of polymer coatings to answer key fundamental questions in cell biology using 2D coatings, and to develop polymer-coated microcarriers that provide an efficient, well-defined platform for cell biomanufacturing. These key fundamental goals include: i) understanding the influence of local and global coating compositions on cellular behaviors; ii) understanding the role of substrate-mediated growth factor (GF) sequestering on GF-dependent cell expansion; and iii) exploring the influence of controlled, divalent presentation of receptor-binding peptides on receptor activation and associated cell behavior. These studies will result in the design of an efficient copolymer coating and ligation chemistry to present receptor-coreceptor clusters with controlled spacing to study their influence on GF signaling and associated cell behaviors. To quantitatively characterize the functionality of the surface, this research will develop Time of Flight-Secondary Ion Mass Spectrometry (TOF-SIMS) in conjunction with X-ray photoelectron spectroscopy (XPS), and assess the local and global heterogeneities in surface composition. The outcome of this research will be the synthesis of chemically defined microcarriers via a copolymer chemistry that is customizable to different cell types and culture media with relevant bio-inspired peptides that regulate cell adhesion and GF sequestering. We envision a new class of microcarriers that use customizable polymer coatings, lower the currently intractable cost of media formulations, and achieve efficient, xeno-free expansion of functional therapeutic cells. The ability to tailor microcarriers to support adhesion and receptor activation of specific cell types without using complex and expensive media would be transformative in biomanufacturing, which is a rapidly growing segment of health care in cell and tissue therapy.

第 1 部分：非技术性在过去三十年中，各种基础科学发现已经确定了可用于医学研究和治疗的细胞。然而，为了得到有效使用，这些电池必须高效、可控和可重复地制造。培养和扩增细胞，同时增强其所需功能对于生物制造至关重要，对于最终利用细胞来理解和治疗人类疾病也至关重要。该提案重点关注两种重要细胞类型的生物制造，即已用于 500 多项人体临床试验的人类间充质干细胞 (hMSC) 和形成人类血管的内皮细胞 (EC)。在体外更好地理解和控制 hMSC 和 EC 行为（例如粘附、增殖和分化）的调节可能有助于它们的有效制造并提高细胞疗法的成功率。该研究项目将通过开发生物材料来模仿天然细胞外基质（ECM）的部分，从而了解细胞行为，并更有效地制造治疗细胞。这些聚合物涂层形式的生物材料将有助于回答细胞生物学中的关键基本问题，并为细胞生物制造提供一个高效、明确的平台。由此产生的用于 3D 细胞扩增的聚合物涂层“微载体”可以交付给细胞生物学家和生物工程师，然后他们可以定制它们以探索关键的生物学问题。通过这项研究开发的干细胞和组织工程实践展览将由 PI 和研究生在全国范围内传播。该研究计划将通过与研究生导师合作，激发本科生并让其参与聚合物科学和工程研究。第 2 部分：技术摘要本研究计划将研究聚合物涂层的合成，以利用 2D 涂层回答细胞生物学中的关键基本问题，并开发聚合物涂层微载体，为细胞生物制造提供高效、明确的平台。这些关键的基本目标包括： i) 了解局部和整体涂层成分对细胞行为的影响； ii) 了解底物介导的生长因子 (GF) 隔离对 GF 依赖性细胞扩增的作用； iii) 探索受体结合肽的受控二价呈现对受体激活和相关细胞行为的影响。这些研究将设计出有效的共聚物涂层和连接化学，以呈现具有受控间距的受体-共受体簇，以研究它们对 GF 信号传导和相关细胞行为的影响。为了定量表征表面的功能，本研究将结合 X 射线光电子能谱 (XPS) 开发飞行时间二次离子质谱 (TOF-SIMS)，并评估表面成分的局部和全局异质性。这项研究的成果将是通过共聚物化学合成化学成分确定的微载体，该共聚物化学可根据不同的细胞类型和培养基进行定制，并含有调节细胞粘附和 GF 隔离的相关仿生肽。我们设想一种新型微载体，它使用可定制的聚合物涂层，降低目前棘手的培养基配方成本，并实现功能性治疗细胞的高效、无异源扩增。在不使用复杂且昂贵的培养基的情况下定制微载体以支持特定细胞类型的粘附和受体激活的能力将给生物制造带来变革，生物制造是细胞和组织治疗中快速增长的医疗保健领域。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Customized hydrogel substrates for serum-free expansion of functional hMSCs.

用于功能性 hMSC 的无血清扩增的定制水凝胶基质。

DOI：
10.1039/d0bm00540a
发表时间：
2020
期刊：
RSC biomolecular sciences
影响因子：
0
作者：
Le, Nhi T;Liu, Leona Tianran;Johnston, James;Krutty, John D;Templeton, Kayla M;Harms, Victoria;Dias, Andrew;Le, Hau;Gopalan, Padma;Murphy, William M.
通讯作者：
Murphy, William M.

Polymer-Coated Magnetic Microspheres Conjugated with Growth Factor Receptor Binding Peptides Enable Cell Sorting