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Three-Dimensional Cardiac Gel Bioreactor for Functioning Cardiomyocyte Induction

用于诱导功能性心肌细胞的三维心脏凝胶生物反应器

基本信息

批准号：
7821360
负责人：
KIMIMASA TOBITA
金额：
$ 18.94万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7821360
关键词：
3-Dimensional Abbreviations Actins Adipose tissue Age Allogenic Animals Autologous Biopsy Bioreactors Bone Marrow Calcium ion Cardiac Cardiac Myocytes Cardiac Myosins Cardiomyoplasty Cells Clinical Trials Connexin 43 Digestion Electric Stimulation Embryo Engineering Evaluation Gel Gene Proteins Goals Heart Heart Diseases Hematopoietic Human Hypoxia Implant In Vitro Investigation Mechanics Methods Molecular Genetics Muscle Muscle satellite cell Myocardial Myocardial tissue Myocardium Myosin Heavy Chains Natural regeneration Nerve Patients Pericytes Property Rattus Reporting Research Design Role Skeletal Muscle Skeletal Myoblasts Smooth Muscle Myocytes Source Specimen Stem cells Stretching System Tendon structure Testing Tissues Transplantation Tretinoin Troponin I Troponin T Undifferentiated Work bone embryonic stem cell fetal improved injured progenitor public health relevance reconstitution repaired stem stem cell population tissue culture

项目摘要

DESCRIPTION (provided by applicant): The limited capacity of the injured myocardium to reactivate cardiomyocyte (CM) proliferation is a major barrier to myocardial regeneration. Stem cell derived CMs are considered as an alternative cell source to replace damaged CMs. Various types of stem cells are under investigation of their potential for CM differentiation. Skeletal muscle derived stem cells (MDSCs) are a somatic stem cell population obtained from skeletal muscle specimens. MDSCs have been shown to differentiate along multiple lineages including skeletal muscle, bone, tendon, nerve, endothelial, hematopoietic, and smooth muscle cells. MDSCs are easily expanded in vitro and can be transplanted as autologous grafts. Previous studies, including our own work, have shown that MDSCs have potential for cardiac repair and can differentiate into CMs. We have recently succeeded in the induction of functioning CM from cultured rat MDSCs using the strategy of combined MDSC-aggregate sphere formation and MDSC-sphere-3-dimensional cardiac gel bioreactor (MDSC-sphere-3DGB). MDSC-sphere-3DGB is scalable and the MDSC-derived CMs are easily extracted by enzymatic digestion of the construct. MDSC-sphere-3DGB also allows the direct evaluation of MDSC-derived CM contractile properties as an engineered cardiac tissue. Thus, the aim of the proposal is to optimize a tissue culture method to obtain functioning CMs differentiated from human MDSCs. Specific Aim: Define the optimal condition of human MDSC-sphere-3DGB for functioning CM induction. We will test 4 hypotheses, 1). Human MDSCs differentiate into functioning CMs using a MDSC-sphere-3DGB system; 2). Additive retinoic acid treatment in MDSC-sphere-3DGB further increases functioning CM differentiation; 3). Cyclic mechanical stretch stimulation increases human MDSC-derived CM proliferation and contractile properties similar to the immature fetal type CMs; 4). Cyclic electric stimulation increases human MDSC-derived CM maturation and improves tolerance to the hypoxic conditions. Significance: The goal of the present proposal is to establish a paradigm using a MDSC-sphere-3DGB system to define the differentiation of the functioning human MDSC-derived CMs. These studies support our long term goal of implementing autologous cellular cardiomyoplasty to repair congenital and acquired heart diseases. PUBLIC HEALTH RELEVANCE: We have recently succeeded in generating workable heart muscle cell from rat skeletal muscle derived stem cells using a 3-dimensional tissue culture system (bioreactor). The skeletal muscle derived stem cell is relatively easily obtained from a skeletal muscle biopsy of heart disease patients and the induced heart muscle cells from the skeletal muscle stem cells can be transplanted to the same patients to cure the damaged heart. This proposal is to investigate the optimal conditions of HUMAN skeletal muscle derived stem cell-derived heart muscle cells to cure congenital and acquired heart diseases.

描述（由申请人提供）：受损心肌重新激活心肌细胞（CM）增殖的能力有限，是心肌再生的主要障碍。干细胞衍生的 CM 被认为是替代受损 CM 的替代细胞来源。正在研究各种类型的干细胞的 CM 分化潜力。骨骼肌衍生干细胞（MDSC）是从骨骼肌标本中获得的成体干细胞群。 MDSC 已被证明可沿多个谱系分化，包括骨骼肌、骨、肌腱、神经、内皮、造血和平滑肌细胞。 MDSC 很容易在体外扩增，并且可以作为自体移植物进行移植。之前的研究，包括我们自己的工作，已经表明 MDSC 具有心脏修复潜力，并且可以分化为 CM。我们最近利用 MDSC-聚集球形成和 MDSC-sphere-3 维心脏凝胶生物反应器 (MDSC-sphere-3DGB) 相结合的策略，成功地从培养的大鼠 MDSC 中诱导了功能性 CM。 MDSC-sphere-3DGB 具有可扩展性，并且可以通过酶消化构建体轻松提取 MDSC 衍生的 CM。 MDSC-sphere-3DGB 还允许直接评估 MDSC 衍生的 CM 作为工程心脏组织的收缩特性。因此，该提案的目的是优化组织培养方法以获得从人类 MDSC 分化而来的功能性 CM。具体目标：定义人类 MDSC-sphere-3DGB 用于功能 CM 诱导的最佳条件。我们将测试 4 个假设，1)。使用 MDSC-sphere-3DGB 系统，人类 MDSC 分化为功能性 CM； 2）。 MDSC-sphere-3DGB 中的附加视黄酸处理进一步增加了功能性 CM 分化； 3）。循环机械拉伸刺激可增加人 MDSC 衍生的 CM 增殖和收缩特性，类似于未成熟胎儿型 CM； 4).循环电刺激可促进人 MDSC 衍生的 CM 成熟并提高对缺氧条件的耐受性。意义：本提案的目标是建立一个使用 MDSC-sphere-3DGB 系统的范例来定义功能性人类 MDSC 衍生 CM 的分化。这些研究支持我们实施自体细胞心肌成形术来修复先天性和后天性心脏病的长期目标。公共健康相关性：我们最近使用 3 维组织培养系统（生物反应器）成功地从大鼠骨骼肌来源的干细胞中产生了可用的心肌细胞。骨骼肌来源的干细胞相对容易从心脏病患者的骨骼肌活检中获得，并且来自骨骼肌干细胞的诱导心肌细胞可以移植到同一患者体内以治愈受损的心脏。该提案旨在研究人类骨骼肌干细胞衍生的心肌细胞治疗先天性和后天性心脏病的最佳条件。