Induction of neuro-protective/regenerative genes in stem cells infiltrating post-ischemic brain tissue - PubMed (original) (raw)
Induction of neuro-protective/regenerative genes in stem cells infiltrating post-ischemic brain tissue
Gokhan Yilmaz et al. Exp Transl Stroke Med. 2010.
Abstract
Background: Although the therapeutic potential of bone marrow-derived stromal stem cells (BMSC) has been demonstrated in different experimental models of ischemic stroke, it remains unclear how stem cells (SC) induce neuroprotection following stroke. In this study, we describe a novel method for isolating BMSC that infiltrate postischemic brain tissue and use this method to identify the genes that are persistently activated or depressed in BMSC that infiltrate brain tissue following ischemic stroke.
Methods: Ischemic strokes were induced in C57BL/6 mice by middle cerebral artery occlusion for 1 h, followed by reperfusion. BMSC were isolated from H-2 Kb-tsA58 (immortomouse) mice, and were administered (i.v.) 24 h after reperfusion. At the peak of therapeutic improvement (14 days after the ischemic insult), infarcted brain tissue was isolated, and the BMSC were isolated by culturing at 33 degrees C. Microarray analysis and RT-PCR were performed to compare differential gene expression between naïve and infiltrating BMSC populations.
Results: Z-scoring revealed dramatic differences in the expression of extracellular genes between naïve and infiltrating BMSC. Pair-wise analysis detected 80 extracellular factor genes that were up-regulated (>/= 2 fold, P < 0.05, Benjamini-Hochberg correction) between naïve and infiltrated BMSC. Although several anticipated neuroregenerative, nerve guidance and angiogenic factor (e.g., bFGF, bone morphogenetic protein, angiopoietins, neural growth factor) genes exhibited an increased expression, a remarkable induction of genes for nerve guidance survival (e.g., cytokine receptor-like factor 1, glypican 1, Dickkopf homolog 2, osteopontin) was also noted.
Conclusions: BMSC infiltrating the post-ischemic brain exhibit persistent epigenetic changes in gene expression for numerous extracellular genes, compared to their naïve counterparts. These genes are relevant to the neuroprotection, regeneration and angiogenesis previously described following stem cell therapy in animal models of ischemic stroke.
Figures
Figure 1
Neurological scores of sham, stroke, stroke +WT BMSC and stroke + Large T BMSC groups over a two week period after stroke or sham surgery. * represents significant difference from sham group (p < 0.05), # represents significant difference from stroke group (P < 0.05), one-way ANOVA with Tukey's post-hoc test.
Figure 2
Large T BMSC isolated from ischemic brain. A. Nuclear DAPI Staining, B. Large T immunostaining of the same sample.
Figure 3
Volcano graph of gene expression changes in BMSC isolated from ischemic brain (Group1) versus naïve BMSC (Group2).
Figure 4
Fold-change in gene expression detected by RT-PCR versus microarray.
Figure 5
Z-scores of gene ontology groups that changed in BMSC isolated from ischemic brain.
Figure 6
Changes in KEGG pathways in BMSC isolated from ischemic brain. Changes filtered for parameters Z-score > 2 and 10 or more changed genes in related group.
Figure 7
Axon guidance KEGG pathway affected in BMSC isolated from ischemic brain. Genes affected represented in red.
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