Department of Neuroregeneration, Netherlands Institute for Neurosciences, Amsterdam, The Netherlands Department of Neurosurgery, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
Restor Neurol Neurosci. 2011;29(3):187-201. doi: 10.3233/RNN-2011-0591.
The assessment of the capacity of bone marrow stromal cells (BMSC) to repair the nervous system using gene expression profiling. The evaluation of effects of long-term culturing on the gene expression profile of BMSC.
Fourty four k whole genome rat microarrays were used to study gene expression of cultured BMSC at passage (P)3 and to compare expression profiles between P3 and P14 BMSC. Quantitative PCR was employed to validate the microarray results.
P3 BMSC expressed genes involved in neural developmental events such as glial differentiation, neuron proliferation, and neurite formation. They also express genes encoding for growth factors and for proteins involved in growth factor signaling. A total of 6687 genes were co-expressed in P3 and P14 BMSC. Of these co-expressed genes, 3% (202 genes) was differentially expressed with 159 genes higher in P3 BMSC and 43 genes higher in P14 BMSC. The gene expression patterns were independently validated using quantitative PCR. Functional data mining by Gene Ontology (GO)-analysis revealed that 85/159 and 22/43 genes were annotated in the GO database. In P3 BMSC, 53 GO-classes were overrepresented with several involved in organ development, cell proliferation, and neural repair. In P14 BMSC, three GO-classes were overrepresented with one involved in organ development.
Our gene profiling results suggested a decreased plasticity and repair aptitude of long-term cultured BMSC. Our data indicated the use of early passage BMSC for neural repair approaches.
利用基因表达谱评估骨髓基质细胞 (BMSC) 修复神经系统的能力。评估长期培养对 BMSC 基因表达谱的影响。
使用 44k 全基因组大鼠微阵列研究培养的 BMSC 在第 3 代(P3)的基因表达,并比较 P3 和 P14 BMSC 之间的表达谱。采用定量 PCR 验证微阵列结果。
P3 BMSC 表达了参与神经发育事件的基因,如神经胶质分化、神经元增殖和神经突形成。它们还表达编码生长因子和参与生长因子信号的蛋白质的基因。在 P3 和 P14 BMSC 中共表达了 6687 个基因。在这些共表达的基因中,有 3%(202 个基因)在 P3 BMSC 中差异表达,其中 159 个基因上调,43 个基因下调。使用定量 PCR 独立验证了基因表达模式。通过基因本体论(GO)分析的功能数据挖掘显示,在 GO 数据库中有 85/159 和 22/43 个基因被注释。在 P3 BMSC 中,有 53 个 GO 类被过度表达,其中一些与器官发育、细胞增殖和神经修复有关。在 P14 BMSC 中,有三个 GO 类被过度表达,其中一个与器官发育有关。
我们的基因谱分析结果表明,长期培养的 BMSC 的可塑性和修复能力降低。我们的数据表明,早期传代的 BMSC 可用于神经修复方法。