Arnhold Stefan, Klein Helmut, Klinz Franz-Josef, Absenger Yvonne, Schmidt Annette, Schinköthe Timo, Brixius Klara, Kozlowski Jolanta, Desai Biren, Bloch Wilhelm, Addicks Klaus
Department of Anatomy I, University of Cologne, Josef-Stelzmannstr. 9, D-50931 Köln, Germany.
Eur J Cell Biol. 2006 Jun;85(6):551-65. doi: 10.1016/j.ejcb.2006.01.015. Epub 2006 May 2.
Because the neural differentiation capacity of bone marrow stromal cells (BMSCs) is still a matter of controversial debate, we performed a thorough investigation into the differentiation capacity of human BMSCs and examined their therapeutic potency. BMSCs were isolated from the femur and kept in cell cultures with various cultivation protocols being applied. In standard culture conditions using a fetal calf serum-enriched medium, while not exhibiting a neural phenotype, the majority of cells expressed a variety of neuronal marker proteins as well as the astrocyte marker GFAP. Only a minority of stem cells expressed nestin, a marker for neural precursor cells. Cultivation in serum-free medium supplemented with specific growth factors resulted in a markedly higher percentage of nestin-positive cells. To establish the therapeutic potency of bone marrow-derived cells, the synthesis of neurotrophic factors such as NGF, BDNF and GDNF was analyzed under non-stimulating standard culture conditions as well as after a neural selection procedure. The therapeutic potency of BMSCs was further examined with regard to their migratory potential in vitro and after transplantation in vivo. After stereotactic engraftment into the lateral ventricle of adult rats, mesenchymal stem cells were seen to adhere to the ependymocytes and cells of the choroids plexus. Afterwards grafted cells passed through the ependymal barrier, locating in the subventricular space. Their BMSCs took up a close host graft interaction without any degenerative influence on the host cells. Furthermore, there was morphological as well as immunohistochemical evidence for a transdifferentiation within the host tissue. In addition, BMSCs could be efficiently transduced using a third-generation adenoviral vector, indicating their potential feasibility for a gene-therapeutic option.
由于骨髓基质细胞(BMSCs)的神经分化能力仍存在争议,我们对人BMSCs的分化能力进行了深入研究,并检测了它们的治疗潜力。从股骨中分离出BMSCs,并采用不同的培养方案进行细胞培养。在使用富含胎牛血清的培养基的标准培养条件下,大多数细胞虽未表现出神经表型,但表达了多种神经元标记蛋白以及星形胶质细胞标记物GFAP。只有少数干细胞表达神经前体细胞标记物巢蛋白。在添加特定生长因子的无血清培养基中培养,巢蛋白阳性细胞的比例显著更高。为确定骨髓来源细胞的治疗潜力,在非刺激的标准培养条件下以及经过神经选择程序后,分析了神经营养因子如NGF、BDNF和GDNF的合成。还进一步检测了BMSCs在体外和体内移植后的迁移潜力。在立体定向植入成年大鼠侧脑室后,间充质干细胞可黏附于室管膜细胞和脉络丛细胞。随后移植细胞穿过室管膜屏障,定位在脑室下区。它们的BMSCs与宿主移植物紧密相互作用,对宿主细胞无任何退行性影响。此外,有形态学和免疫组织化学证据表明宿主组织内发生了转分化。此外,使用第三代腺病毒载体可有效转导BMSCs,表明它们在基因治疗方面具有潜在的可行性。