Mechanisms of multipotency of adult stem cells derived from bone marrow stroma and peripheral blood and their application for regenerative medicine

为了实现理想的生殖医学，确保干细胞的稳定供应是极其重要的，并且在使用ES细胞的情况下，使用患者自身的体细胞进行器官和组织的生殖有望实现理想的治疗，而没有许多限制。因此，本研究以骨髓基质细胞为研究对象，探讨其在生殖医学中的应用前景。首先，从小鼠骨髓中分离培养了多株骨髓基质细胞。我们成功地建立了一种新的方法，通过与各种生长因子、维生素和细胞外基质一起培养，诱导这些细胞分化为成骨细胞、软骨细胞、肌肉细胞和脂肪细胞等中胚层细胞、神经细胞等外胚层细胞和肝细胞等内胚层细胞。有趣的是，在这些诱导条件下，几乎可以100%诱导神经细胞和肝细胞。这些结果表明 关于我们 然后，将绿色荧光蛋白（GFP）基因导入这些体外显示多能性的骨髓基质细胞系，建立稳定表达GFP的细胞系。在GFP基因导入过程中，一些细胞系表现出不同程度的分化。在进一步的体外检查后，将多能培养细胞接种到免疫缺陷小鼠的皮下组织、肌肉、脑、肝、人工制造的缺乏骨和软骨的区域以及脊髓损伤部位，并随时间收集包含接种部位的组织和器官，以组织学检查移植细胞的定位和分布。本研究中新建立的一些小鼠骨髓基质细胞系显示出完全的多能性，而另一些显示出有限的多能性。因此，通过比较这些不同表型中的基因表达，正在探索参与维持未分化状态的基因。少

For realization of ideal reproductive medicine, it is extremely important to secure the stable supply of stem cells, and reproduction of organs and tissues using patients' own somatic cells is expected to realize ideal therapy without a number of restrictions in cases of using ES cells. From this point of view, we here focused on multipotency of bone marrow stromal cells and investigated the potential application for reproductive medicine.First, several lines of bone marrow stromal cells were established from mouse bone marrow. We successfully established a new method to induce differentiation of these cells into mesodermal cells such as osteoblasts, cartilage cells, muscle cells, and adipocytes, ectodermal cells such as nerve cells, and endodermal cells such as hepatocytes by culture with various growth factors, vitamins, and extracellular matrix. Intriguingly, it was possible to induce nerve cells and hepatocytes at almost 100% under these induction conditions. These results indicate … More d that multipotent adult stem cells were present in bone marrow stromal cells.Next, the green fluorescent protein (GFP) gene was introduced to these bone marrow stromal cell lines exhibiting multipotency in vitro and stable cell lines expressing GFP were established. During the GFP gene introduction, some cell lines showed different degrees of differentiation. After additional in vitro examination, multipotent culture cells were inoculated in the subcutaneous tissue, muscles, brain, liver, artificially created areas lacking bones and cartilages, and spinal injury sites in immunodeficient mice, and tissues and organs containing the inoculation site were collected over time to histologically examine the localization and distribution of transplanted cells.As a result, some mouse bone marrow stromal cell lines newly established in this study showed complete multipotency while others revealed limited multipotency. Therefore, exploration of genes involved in maintenance of an undifferentiated state is underway by comparing gene expression in these different phenotypes. Less