IN VIVO DIFFERENTIATION AND MIGRATION PROPERTIES OF STEM CELLS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 越来越多的证据表明，从骨髓或脂肪组织中分离的间充质干细胞（MSC）是多能细胞，可以在CNS中移植和分化。如果开发用于细胞疗法，MSC可以提供优势，因为这些细胞相对容易从小体积的脂肪抽吸物或骨髓抽吸物中分离，并且可以容易地扩增以提供足够数量的细胞用于移植。在这项研究中，进行了一系列的中枢神经系统移植实验，以表征人类骨髓间充质干细胞的植入，迁移，归巢和分化模式。将人骨髓源性间充质干细胞（BMSCs）和脂肪组织源性间充质干细胞（ASCs）经立体定位注射入NIH Ⅲ免疫缺陷小鼠的侧脑室。注射后15天和30天通过免疫组织化学和PCR分析监测MSC的命运。MSCs从注射部位开始沿脑室区沿着向嘴侧和尾侧迁移。细胞分布于整个脑室区的吻尾侧范围。在整个心室的室管膜中也检测到许多MSC。大量供体细胞沿脊髓长度分布沿着。在注射后15天，在注射ASC的动物中检测到的MSC的频率高于BMSC。供体细胞分布于大脑的两个脑叶，在大脑皮质和不同的区域，如半月小叶，旁绒球，后上，裂区，内外颗粒层中检测到大量的细胞。详细的分析显示，MSC主要见于脑血管周围空间。在注射ASC的动物中，可检测到的细胞总数在植入后30天显著高于15天，尽管分布模式没有改变。30 d组脊髓内移植的骨髓间充质干细胞数量明显增多。在两个时间点，BMSC广泛分布于整个脑中。骨髓间充质干细胞在移植后15天的分布模式与ASCs相似。在30天时，大部分细胞被发现在脑的腹侧区域中，在基底动脉的直接周边中检测到许多细胞。这些研究的结果表明，MSCS通过脑脊液有效地迁移到整个CNS，因为细胞的检测模式与脑中脑脊液的流动平行。在移植后30天，发现许多MSC对一些特异性神经元抗体如NSE和MAP-2具有反应性，表明移植细胞的分化。这些研究的结果表明，将两种类型的MSC移植到脑室中导致MSC在纹状体、皮质和小脑区域以及脊髓内的整个脑中的迁移。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There is increasing evidence that mesenchymal stem cells (MSCs) isolated from the bone marrow or adipose tissue are multipotent cells that can engraft and differentiate in the CNS. MSCs may offer advantages if developed for cell therapy because these cells are relatively easy to isolate from small volume of lipoaspirate or of bone marrow aspirate and can be readily expanded to provide a sufficient number of cells for transplantation. In this study, a series of CNS transplantation experiments were performed to characterize the engraftment, migration, homing, and differentiation patterns of human MSCs. Cell lines generated from human bone marrow derived MSCs (BMSCs) and adipose tissue derived MSC (ASCs) were injected into lateral ventricle of brain of NIHIII immune deficient mice by stereotaxic injection. The fate of MSCs was monitored by immunohistochemistry and PCR analyses 15 and 30 days post-injection. MSCs migrated along ventricular area both rostrally and caudally from the injection site. Cells were distributed throughout the entire rostro-caudal extent of ventricular zone. Many MSCs also were detected lining the ependyma throughout the ventricle. A significant number of donor cells were distributed along the length of the spinal cord. A higher frequency of MSCs was detected in animals injected with ASCs than BMSCs at 15 days following injection. Donor cells were dispersed into both lobes of the brain, numerous cells were detected in the cortex and different areas of cerebrum such as semilunar lobules, paraflocculus, postsuperior, fissure areas, and external and internal granular layers. Detailed analysis revealed that MSCs were predominantly seen in perivascular space in the brain. In animals injected with ASCs, the total number of detectable cells was significantly higher in 30 days post-engraftment than 15 days, though the distribution pattern was unchanged. A significantly higher number of engrafted BMSCs were detected in the spinal cord of 30 days group. BMSCs were widely distributed throughout the brain at both time points. The distribution pattern of BMSCs at 15 days post engraftment was similar to ASCs. At 30 days, the majority of cells were found in the ventral areas of brain, with many detected in the immediate periphery of basilary artery. The results form these studies suggest that the MSCS effectively migrate throughout the CNS via the cerebrospinal fluid, as the pattern of detection of cells parallels the flow of cerebrospinal fluid in the brain. Numerous MSCs were found to be reactive to some specific neuronal antibodies such as NSE and MAP-2 at 30 days following engraftment indicating the differentiation of the grafted cells. The results of these studies demonstrated that transplantation of both types of MSCs into the ventricle resulted in the migration of MSCs throughout the brain within striatal, cortical, and cerebellar regions as well as and spinal cord.