Can Stromal Cells Differentiate into Oligodendrocytes?

描述（由申请人提供）：在多发性硬化症中， 神经症状归因于脱髓鞘和轴突损伤。我们 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 是利用细胞移植来帮助修复过程。基于 我们的初步结果，我们假设骨髓来源的人， 间充质基质细胞（hMSC）可以分化为少突胶质细胞， 因此可以是用于这种治疗的细胞来源。 近期目标是：目标1。以产生纯化的少突胶质细胞 从培养的hMSCs中。将hMSC铺在有利于生长的表面上， 少突胶质细胞发育，然后用已知的生长因子处理， 从胚胎或神经干细胞产生少突胶质细胞。所得 细胞群将被表征以确定细胞的比例 少突胶质细胞、星形胶质细胞、神经元和小胶质细胞谱系。我们将 确定生长因子和培养条件对产生 少突胶质细胞前体，这样我们就可以最大限度地增加 用于移植的少突胶质细胞祖细胞。净化方法准备就绪 在我们的实验室中用于大鼠和小鼠细胞。目标二。到 研究神经调节蛋白和delta/jagged在分化中的重要性 将hMSC 5转化为少突胶质细胞和星形胶质细胞谱系。细胞会暴露在 GGF 2和jagged/delta以及OL和星形胶质细胞标志物的表达将被检测。 通过免疫细胞化学方法评估。确认观察结果 锯齿状/δ，hMSC将被转染组成型激活的 notch 1受体，这些细胞将研究OL的表达， 星形胶质细胞标记目标3。目的探讨hMSC细胞体外培养的可行性 保留区分为OL的能力的行。为了生成这样的线条 人端粒酶催化亚单位v-myc和H-ras将用于 感染hMSC。在选择细胞并确定细胞的表达后， 外源基因，细胞系分化为OL的能力将被 测试.在这三个目标中，OL分化的细胞培养工作将是 移植研究的支持。 足够数量的细胞或细胞系能够容易地获得， 区分OL应该被认为是解决这个问题的第一步 自体细胞移植作为治疗考虑， 脱髓鞘/髓鞘形成障碍疾病。

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.