Stromal Stem Cells of Human Placenta for the Treatment of Acute Lung Injury

描述(由申请人提供)：需要新的多潜能、高度治疗有效的、患者特有的干细胞来源。胎盘是一种新的潜在来源 胎儿干细胞。涉及胎盘干细胞的治疗用途的研究尚未进行。我们计划通过在美国国立卫生研究院的支持下完成一项研究计划来满足这一需求，该计划将使我们能够开发一种利用选定的胎盘基质细胞治疗急性肺损伤的治疗产品。急性肺损伤是重症患者急性呼吸衰竭的主要原因，需要开发新的治疗模式。该项目将进一步促进干细胞功能知识的进步，并促进使用来自胎盘的干细胞进行再生医学的可能性。我们的假设是，人胎盘的绒毛膜含有大量的间充质干细胞，它们释放出几种有益的旁分泌因子。这一假说是基于我们最近的发现，即人类胎盘是一个造血器官，而胎盘是一个含有大量造血细胞的利基市场。人胎盘绒毛膜间充质中存在与基质细胞和胚胎细胞表型相同的其他细胞。这些细胞在旁分泌信号中高度活跃，并分泌大量的生长因子，如HGF、KGF、转化生长因子-β、成纤维细胞生长因子、GM-CSF和血管生成素。胎盘基质细胞可有效修复受损上皮层的屏障功能。在具体目标1中，我们将检测人胎盘来源的间充质绒毛细胞的多能性及其高旁分泌活性。我们将建立绒毛膜间充质细胞系，并对其增殖和分化潜能进行鉴定。细胞的特征将包括其形态、在培养中增殖的能力、人类胚胎干细胞标志(SSEA-3、OCT-4、TRA-1-60)的表达以及类胚体的形成。采用酶联免疫吸附试验检测细胞旁分泌活性。治疗细胞将从20个胎盘中根据HGF、FGF、TGF-β、GM-CSF和血管生成素的分泌水平而选择。在特定的目标2中，我们将测试选定的胎盘基质干细胞在动物和人类急性肺损伤模型中的潜在治疗价值。细胞将被用于治疗急性肺损伤的小鼠 并对肺纤维化的发展起到促进作用。在体外灌流的人肺制剂中，胎盘细胞在促进受损的肺上皮和内皮修复方面的有效性将被进一步研究。该项目将提供丰富、无争议和廉价的治疗活性基质干细胞来源，在非常短的时间内(2-4年)可用于人类。 与公共卫生相关：迫切需要多能、高度治疗有效的患者专用干细胞的新来源。胎盘是胎儿干细胞的潜在来源。很少有研究涉及用于治疗的胎盘干细胞。我们计划在NIH的支持下开发一项研究计划，使我们能够获得一种基于胎盘基质细胞的治疗产品，用于治疗急性肺损伤等急性疾病。细胞将根据其旁分泌活性从胎盘中挑选出来，并在急性肺损伤的动物模型和体外分离的人肺中进行测试。这项研究将进一步促进知识的进步和利用来自胎盘的干细胞进行再生医学的潜力。该项目完成后，将提供新的丰富、无争议和廉价的具有治疗活性的胎儿干细胞来源，在很短的时间内可用于人类，这将决定其未来的巨大商业潜力。

DESCRIPTION (provided by applicant): There is a need for new sources of multipotent, highly therapeutically effective, patient-specific stem cells. The placenta is a novel potential source of fetal stem cells. Studies of therapeutic use involving placental stem cells have not been conducted. We plan to address this need by accomplishing a research program with support from the NIH that will enable us to develop a therapeutic product utilizing selected placental stromal cells for the treatment of Acute Lung Injury. Acute Lung Injury is a major cause of acute respiratory failure in critically ill patients which requires development of novel modes of therapy This project will further contribute to the advances in knowledge of stem cell function and to the potential for regenerative medicine using stem cells derived from the placenta. Our hypothesis is that the chorion of human placenta contains large numbers of mesenchymal stem cells, which release several beneficial paracrine factors. This hypothesis is based on our recent discoveries that human placenta is a hematopoietic organ and term placenta is a niche of abundant numbers of hematopoietic cells. Other cells, sharing the phenotype of stromal cells and embryonic cells are present in the mesenchyme of human placental chorion. These cells are highly active in paracrine signaling and secrete abundant quantities of growth factors like HGF, KGF, TGF-beta, FGF, GM-CSF, and angiopoietins. Stromal cells from the placenta are effective in restoration of barrier function of injured epithelial layers. In Specific Aim 1, we will test th multipotency of mesenchymal chorionic cells derived from human placenta and their high paracrine activity. We will generate chorionic mesenchymal cell lines and characterize their proliferation and differentiation potential. Cells will be characterized by their morphology, abiliy to propagate in culture, expression of markers of human embryonic stem cells (SSEA-3, Oct-4, TRA-1-60), and embryoid body formation. We will also measure the paracrine activity of cells by ELISA. Cells for therapy will be selected from 20 placentas upon the levels of secretion of HGF, FGF, TGF-beta, GM-CSF, and angiopoietins. In Specific Aim 2, we will test the potential therapeutic value of selected placental stromal stem cells for their efficacy in animal and human lung models of acute lung injury. Cells will be tested for treatment of mice with acute lung injury and for development of lung fibrosis. Placental cells will be further investigated for their effectiveness in facilitation of repair of injured lung epithelium and endothelium in an ex vivo perfused human lung preparation. This project will provide abundant, non-controversial and inexpensive source of therapeutically active stromal stem cells, available for use in humans within a very short period of time (2-4 years). PUBLIC HEALTH RELEVANCE: Novel sources of multipotent, highly therapeutically effective patient-specific stem cells are in great need. The placenta is a potential source of fetal stem cells. Few studies involving placental stem cells for therapeutic use have been conducted. We plan to develop a research program, with support from NIH that will enable us to obtain a therapeutic product based upon use of placental stromal cells for the treatment of acute disorders like Acute Lung Injury. Cells will be selected from placentas upon their paracrine activity and tested in animal models of Acute Lung Injury and in isolated ex vivo human lungs. This study will further contribute to the advances in knowledge and to the potential for regenerative medicine using stem cell derived from the placenta. Upon completion, this project will provide novel abundant, non-controversial and inexpensive source of therapeutically active fetal stem cells, available for use in humans within very short period of time, which will determine its outstanding future commercial potential.