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

描述（由申请人提供）：需要多能、高治疗有效性、患者特异性干细胞的新来源。胎盘是一种新的潜在来源， 胚胎干细胞尚未进行涉及胎盘干细胞的治疗用途的研究。我们计划通过在NIH的支持下完成一项研究计划来满足这一需求，该研究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。急性肺损伤是危重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对干细胞功能的认识，并促进利用胎盘干细胞进行再生医学的潜力。我们的假设是，人胎盘绒毛膜中含有大量的间充质干细胞，这些间充质干细胞释放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，足月胎盘是一个大量造血细胞的小生境。人胎盘绒毛膜的间充质中还存在有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传导中高度活跃，并分泌大量的生长因子，如HGF、KGF、TGF-β、FGF、GM-CSF和血管生成素。来自胎盘的基质细胞可有效恢复受损上皮层的屏障功能。在具体目标1中，我们将检测来自人胎盘的间充质绒毛膜细胞的多能性及其高旁分泌活性。我们将产生绒毛膜间充质细胞系，并表征其增殖和分化潜力。细胞将通过其形态、在培养物中增殖的能力、人胚胎干细胞标志物（SSEA-3、Oct-4、TRA-1-60）的表达和胚状体形成来表征。我们还将通过ELISA测量细胞的旁分泌活性。根据HGF、FGF、TGF-β、GM-CSF和血管生成素的分泌水平，从20个胎盘中选择用于治疗的细胞。在具体目标2中，我们将测试选定的胎盘基质干细胞在急性肺损伤的动物和人肺模型中的有效性的潜在治疗价值。将测试细胞用于治疗患有急性肺损伤的小鼠 以及肺纤维化的发展。将进一步研究胎盘细胞在离体灌注人肺制剂中促进损伤肺上皮和内皮修复的有效性。该项目将提供丰富的，无争议的和廉价的治疗活性基质干细胞来源，可在很短的时间内（2-4年）用于人类。 公共卫生相关性：非常需要多能、高治疗有效性的患者特异性干细胞的新来源。胎盘是胎儿干细胞的潜在来源。很少有研究涉及胎盘干细胞的治疗用途已经进行。我们计划在NIH的支持下开发一个研究项目，这将使我们能够获得基于使用胎盘基质细胞治疗急性疾病（如急性肺损伤）的治疗产品。根据细胞的旁分泌活性从胎盘中选择细胞，并在急性肺损伤的动物模型和离体人肺中进行测试。这项研究将进一步促进知识的进步和利用胎盘干细胞进行再生医学的潜力。完成后，该项目将提供新的丰富，无争议和廉价的治疗活性胎儿干细胞来源，可在很短的时间内用于人类，这将决定其未来的商业潜力。

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.