Can Stromal Cells Differentiate into Oligodendrocytes?

描述(申请人提供)：在多发性硬化症中，持续性 神经症状归因于脱髓鞘和轴突损伤。我们的 长期目标是建立一个科学的治疗基础， 改善这些患者的症状和体征。一种治疗方法 是使用细胞进行移植，以帮助修复过程。基于 我们的初步结果是，我们假设骨髓来源于人类 间充质基质细胞可分化为少突胶质细胞。 因此，这可能是此类疗法的细胞来源。 近期的目标是：目标1.培养纯化的少突胶质细胞 来源于培养中的hMSCs。人骨髓间充质干细胞将在有利于 少突胶质细胞发育，然后用已知的生长因子处理 从胚胎干细胞或神经干细胞产生少突胶质细胞。由此产生的 细胞群的特征将决定细胞的比例 少突胶质细胞、星形胶质细胞、神经元和小胶质细胞谱系。我们会 确定生长因子和培养条件对世代的影响 这样我们就可以最大限度地增加少突胶质细胞的数量和存活率 用于移植的少突胶质祖细胞。纯化方法都准备好了 将在我们实验室的大鼠和小鼠细胞上使用。目标2.目标 研究-中性调节蛋白和Delta/锯齿状蛋白在分化中的重要性 HMSC5分化为少突胶质细胞和星形胶质细胞系。单元格将暴露在 GGF2和锯齿状/增量以及OL和星形细胞标志物的表达 通过免疫细胞化学方法进行评估。用…来证实观察结果 锯齿状/三角区，hMSCs将以组成性激活的方式转染 NOTCH1受体和这些细胞将被研究OL和 星形细胞标记。目的3.探讨体外培养人骨髓间充质干细胞的可行性 保留了分化为OL的能力的线路。要生成这样的行 人端粒酶催化亚基v-myc和H-ras将用于 感染hMSC。在选择细胞后，并确定其表达 外源基因后，细胞系分化为OL的能力将被 测试过。在这三个目标中，细胞培养对OL分化的作用将是 由移植研究提供支持。 容易获得足够数量的细胞或细胞系，能够 要解决这个问题，首先应该考虑将差异化纳入到OL中 将自体细胞移植作为一种治疗考虑 脱髓鞘/髓鞘障碍疾病。

DESCRIPTION (provided by applicant): In Multiple Sclerosis, persistence of neurological symptoms is attributed to the demyelination and axonal injury. Our long term goals are to establish a scientific basis of therapies that will ameliorate the symptoms and signs that these patients. One therapeutic approach is the use of cells for transplantation to aid in the repair process. Based on our preliminary results, we hypothesize that bone marrow derived human mesenchymal stromal cell (hMSC) can differentiate into oligodendrocytes and thus may be source of cells for such therapies. The immediate goals are: Aim 1. To generate purified oligodendroglial cells from hMSCs in culture. hMSCs will be plated on surfaces that favor oligodendrocyte development and then treated with growth factors known to generate oligodendrocytes from embryonic or neural stem cells. The resulting populations of cells will be characterized to determine the proportion of cells of the oligodendrocyte, astrocyte, neuron, and microglial lineages. We will determine the effect of growth factors and culture conditions on generation of oligodendrocyte precursors so that we can maximize the numbers and survival of oligodendrocyte progenitors for transplantation. Purification methods all ready used on rat and mouse cells in our laboratory will be employed. Aim 2. To investigate the importance of -neuregulin and delta/jagged on differentiation of hMSC5 into oligodendrocyte and astrocyte lineages. The cells will be exposed GGF2 and jagged/delta and the expression of OL and astrocytic markers will be assessed by immunocytochemical methods. To confirm observations with jagged/delta, the hMSCs will be transfected with constitutively activated notch1 receptor and these cells will be studied for expression of OL and astrocytic markers. Aim 3. To investigate the feasibity of generating hMSC cell lines that retain the capacity to differentiate into OL. To generate such lines the catalytic subunit of human telomerase, v-myc and H-ras will be used to infect hMSC. After selection of cells, and determining the expression of the exogenous gene, the ability of the cell lines differentiate into OL will be tested. In all three aims, the cell culture work on OL differentiation will be supported by transplantation studies. The easy availability of sufficient number of cells or cell lines capable of differentiating into OL should be considered as a first to address the question of autologous cell transplantation as a therapeutic consideration for demyelinating/dysmyelinating diseases.