Use of umbilical cord as a unique stem cell source for tissue regeneration

使用脐带作为组织再生的独特干细胞来源

• 批准号: 7931307
• 负责人: XIAO-DONG CHEN
• 金额: --
• 依托单位: SOUTH TEXAS VETERANS HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-10-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7931307
• 关键词: Adhesions; Adipocytes; Adult; Aging; Agreement; Alternative Therapies; Animal Model; Blood Vessels; Bone Marrow Cells; Cardiac; Cardiac Myocytes; Cell Lineage; Cell Therapy; Cell-Matrix Junction; Cells; Cessation of life; Chondroblast; Clinical; Data; Development; Disease; Ectoderm; Embryo; Endoderm; Extracellular Matrix; Fatty acid glycerol esters; Gene Expression; Germ Layers; Gland; Goals; Graft Rejection; Health; Heart; Heart Transplantation; Heart failure; Hepatocyte; Human; Immunocompromised Host; In Vitro; Intestines; Marrow; Mesenchymal Stem Cells; Mesoderm; Microarray Analysis; Mononuclear; Mus; Muscle; Myocardial; Myocardial Infarction; Myocardium; Natural regeneration; Nerve; Nerve Fibers; Osteoblasts; Outcome Study; Paper; Plastics; Population; Positioning Attribute; Procedures; Property; Reliance; Reporting; Research; Retrieval; Signal Transduction; Source; Stem Cell Development; Stem cells; Surveys; System; Teratoma; Testing; Therapeutic; Tissues; Transplantation; Umbilical Cord Blood; Umbilical cord structure; Veterans; age related; angiogenesis; base; bone; clinical application; heart function; human embryonic stem cell; implantation; improved; in vivo; innovation; mouse model; novel therapeutic intervention; reconstruction; repaired; self-renewal; tissue regeneration

DESCRIPTION (provided by applicant): Human umbilical cord blood (UCB) contains mesenchymal stem cells (MSCs) that have higher multipotentiality than adult marrow-derived MSCs (BM-MSCs). However, these cells have been difficult to obtain because the number of MSCs in UCB is extremely low (~5 to 30 out of 1 x 108 mononuclear cells). To date, the isolation of MSCs has depended upon their plastic-adhesion capacity. Most extremely immature MSCs in UCB are likely missed because their ability to adhere to plastic is poor. Our previous studies demonstrated that cellular extracellular matrix (ECM) made by bone marrow cells enhanced MSC attachment and proliferation, and retained their stem cell properties (Chen, et al, 2007, JBMR, 22:1943). Using this system, we found that human UCB may contain a large number of MSCs that adhere to the ECM (at least 10,000- to 100,000-fold greater than that previously reported), but not to plastic. More importantly, implantation of UCB-derived MSCs (UCB-MSCs) obtained by ECM adhesion into immunocompromised mice generated 3 embryonic germ layers-derived tissues including bone, muscle, fat, gland, intestine and nerve fibers. Encouraged by these findings, we propose the hypothesis that human UCB contains a large number of embryonic-like stem cells that have the potential to be used for tissue regeneration in general and myocardial reconstruction in particular. To test this hypothesis, the following 3 Specific Aims will be pursued: Specific Aim 1 is to determine the similarities and differences in global gene expression among human embryonic stem cells (hES cells), UCB-MSCs isolated by ECM adhesion versus those isolated by plastic adhesion, and human adult BM-MSCs, using microarray technology. Specific Aim 2 is to determine the ability of human UCB-MSCs obtained by ECM adhesion to selectively differentiate into desired cell lineages originated from 3 embryonic germ layers in vitro under conditions known to induce commitment to a specific cell lineage, including osteoblasts, adipocytes, chondroblasts and cardiomyocytes (mesoderm), nerve (ectoderm) and hepatocytes (endoderm). Specific Aim 3 is to evaluate the capability of transplanted UCB-MSCs obtained by the ECM adhesion to improve heart function after myocardial infarction (MI) using a well-established mouse model. Despite the great developmental potential of hES cells, there appears to be widespread agreement that a less critical source of cellular material would be preferable for the clinical use of tissue regeneration. If the proposed studies confirm that ECM can enhance retrieval of large numbers of embryonic-like stem cells from UCB, the resulting unlimited source of highly functional UCB-derived MSCs would make it feasible to be alternative to hES cells for cell-based clinical application. Specially, the ultimate outcome of these studies may be a highly practical stem cell- based therapy to treat the post-MI veteran. PUBLIC HEALTH RELEVANCE: As the US population is aging, so too is the US veteran population. According to the 2003 National Survey of Veteran Enrollees' Health and Reliance on VA, 47% of veterans were 65 years or older. Myocardial infarction (MI), as an age-related disease, is one of the leading causes of heart failure and death in the US. Currently, the best option for completely restoring cardiac function after a large MI is heart transplantation. However, it is limited by donor availability and transplant rejection. Recently, regeneration of infracted myocardium by injecting stem cells has been proposed as an alternative therapy. If the proposed studies confirm that umbilical cord blood contains a large number of embryonic- like stem cells that can be isolated by ECM adhesion procedures, and that can selectively differentiate into cardiomyocytes in vitro and reconstruct myocardium in vivo after MI, it will facilitate development of novel therapeutic approaches for cell-based regeneration to treat post-MI veteran.

描述（由申请人提供）： 人脐带血 (UCB) 含有间充质干细胞 (MSC)，其多能性高于成人骨髓来源的 MSC (BM-MSC)。然而，这些细胞很难获得，因为 UCB 中的 MSC 数量极低（1 x 108 个单核细胞中约有 5 至 30 个）。迄今为止，间充质干细胞的分离取决于它们的塑料粘附能力。 UCB 中大多数极其不成熟的 MSC 可能会被遗漏，因为它们粘附塑料的能力很差。我们之前的研究表明，由骨髓细胞产生的细胞外基质（ECM）增强了MSC的附着和增殖，并保留了它们的干细胞特性（Chen等人，2007，JBMR，22：1943）。使用该系统，我们发现人类 UCB 可能含有大量粘附在 ECM 上的 MSC（比之前报道的至少大 10,000 至 100,000 倍），但不粘附在塑料上。更重要的是，将通过ECM粘附获得的UCB源间充质干细胞（UCB-MSCs）植入免疫低下小鼠体内，产生了3个胚胎胚层衍生组织，包括骨、肌肉、脂肪、腺体、肠和神经纤维。受这些发现的鼓舞，我们提出了这样的假设：人类脐带血含有大量胚胎样干细胞，这些干细胞有潜力用于一般组织再生，特别是心肌重建。为了检验这一假设，将追求以下 3 个具体目标： 具体目标 1 是使用微阵列技术确定人胚胎干细胞（hES 细胞）、通过 ECM 粘附分离的 UCB-MSC 与通过塑料粘附分离的 UCB-MSC 以及人类成人 BM-MSC 之间全局基因表达的相似性和差异。具体目标 2 是确定通过 ECM 粘附获得的人 UCB-MSC 在已知诱导分化为特定细胞谱系的条件下，在体外选择性分化为源自 3 个胚胎胚层的所需细胞谱系的能力，包括成骨细胞、脂肪细胞、软骨细胞和心肌细胞（中胚层）、神经（外胚层）和肝细胞 （内胚层）。具体目标 3 是使用成熟的小鼠模型评估通过 ECM 粘附获得的移植 UCB-MSC 改善心肌梗死 (MI) 后心脏功能的能力。 尽管 hES 细胞具有巨大的发展潜力，但人们似乎普遍认为，不太重要的细胞材料来源更适合组织再生的临床应用。如果拟议的研究证实 ECM 可以增强从 UCB 中检索大量胚胎样干细胞，那么由此产生的高功能 UCB 衍生的 MSC 的无限来源将使其可以替代 hES 细胞用于基于细胞的临床应用。特别是，这些研究的最终结果可能是一种高度实用的基于干细胞的疗法，用于治疗心梗后退伍军人。 公共卫生相关性： 随着美国人口老龄化，美国退伍军人人口也在老龄化。根据 2003 年全国退伍军人登记者健康状况和对 VA 依赖程度的调查，47% 的退伍军人年龄在 65 岁或以上。心肌梗塞（MI）作为一种与年龄相关的疾病，是美国心力衰竭和死亡的主要原因之一。目前，大心肌梗死后完全恢复心脏功能的最佳选择是心脏移植。然而，它受到供体可用性和移植排斥的限制。最近，有人提出通过注射干细胞来再生梗塞心肌作为替代疗法。如果拟议的研究证实脐带血含有大量可以通过ECM粘附程序分离的胚胎样干细胞，并且可以在体外选择性分化为心肌细胞并在MI后体内重建心肌，那么将有助于开发基于细胞的再生新治疗方法来治疗MI后退伍军人。