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Multimodal Approaches for Regenerative Stroke Therapies: Combination of Granulocyte Colony-Stimulating Factor with Bone Marrow Mesenchymal Stem Cells is Not Superior to G-CSF Alone.

Balseanu AT, Buga AM, Catalin B, Wagner DC, Boltze J, Zagrean AM, Reymann K, Schaebitz W, Popa-Wagner A - Front Aging Neurosci (2014)

Bottom Line: Functional recovery was tested during the entire post-stroke survival period of 56 days.The combination therapy also led to robust angiogenesis in the formerly infarct core and beyond in the "islet of regeneration." However, G-CSF + BM MSCs may not impact at all on the spatial reference-memory task or infarct volume and therefore did not further improve the post-stroke recovery.We suggest that in a real clinical practice involving older post-stroke patients, successful regenerative therapies would have to be carried out for a much longer time.

View Article: PubMed Central - PubMed

Affiliation: Center of Clinical and Experimental Medicine, University of Medicine and Pharmacy of Craiova , Craiova , Romania.

ABSTRACT
Attractive therapeutic strategies to enhance post-stroke recovery of aged brains include methods of cellular therapy that can enhance the endogenous restorative mechanisms of the injured brain. Since stroke afflicts mostly the elderly, it is highly desirable to test the efficacy of cell therapy in the microenvironment of aged brains that is generally refractory to regeneration. In particular, stem cells from the bone marrow allow an autologous transplantation approach that can be translated in the near future to the clinical practice. Such a bone marrow-derived therapy includes the grafting of stem cells as well as the delayed induction of endogenous stem cell mobilization and homing by the stem cell mobilizer granulocyte colony-stimulating factor (G-CSF). We tested the hypothesis that grafting of bone marrow-derived pre-differentiated mesenchymal cells (BM-MSCs) in G-CSF-treated animals improves the long-term functional outcome in aged rodents. To this end, G-CSF alone (50 μg/kg) or in combination with a single dose (10(6) cells) of rat BM MSCs was administered intravenously to Sprague-Dawley rats at 6 h after transient occlusion (90 min) of the middle cerebral artery. Infarct volume was measured by magnetic resonance imaging at 3 and 48 days post-stroke and additionally by immunhistochemistry at day 56. Functional recovery was tested during the entire post-stroke survival period of 56 days. Daily treatment for post-stroke aged rats with G-CSF led to a robust and consistent improvement of neurological function after 28 days. The combination therapy also led to robust angiogenesis in the formerly infarct core and beyond in the "islet of regeneration." However, G-CSF + BM MSCs may not impact at all on the spatial reference-memory task or infarct volume and therefore did not further improve the post-stroke recovery. We suggest that in a real clinical practice involving older post-stroke patients, successful regenerative therapies would have to be carried out for a much longer time.

No MeSH data available.


Related in: MedlinePlus

Post-stroke neurogenesis and angiogenesis. At 8 weeks post-stroke, none of the DCX+ cells in the SVZ of control animals co-localized with BrdU-labeled nuclei. Instead, the BrdU-positive nuclei were distributed mainly in the “pinwheel” architecture of the ventricular epithelium (A). The DCX+ cells occupied an adjacent, distinct position [(A), arrows]. Some of the DCX+ migrated away from the ventricular wall (B). We noted vigorous neurogenesis with many DCX+ (arrows) co-localizing with BrdU nuclei in the G-CSF-treated animals [(C); arrowheads] and animals treated with G-CSF + BM MSC [(D), arrows]. (E–G) Post-stroke angiogenesis. In regions adjacent to the infarct scar, we found numerous BrdU+ nuclei in the endothelium of newly formed blood vessels in the formerly infarct core [(E), green]. The border to the healthy brain region was abruptly demarcated to the left by NeuN-positive nuclei [(E), red]. Beyond the formerly infarct core, we noted vigorous sprouting angiogenesis as evidenced by RECA/BrdU double positive blood vessels [(F), violet] as well as numerous BrdU+ nuclei in the newly formed endothelium [(F), blue] and reconstruction of the basal lamina [(F), green] during the resolution phase of angiogenesis. By number of laminin/BrdU co-localizations, the density of the newly formed blood vessels was significantly higher (threefold, p = 0.01) in the brains of animals treated with the combination G-CSF + BM MSC as compared to controls and G-CSF alone (G). Cc, corpus callosum; IC, infarct core; IR, islet of regeneration; LV, lateral ventricle; PI, periinfract.
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Figure 5: Post-stroke neurogenesis and angiogenesis. At 8 weeks post-stroke, none of the DCX+ cells in the SVZ of control animals co-localized with BrdU-labeled nuclei. Instead, the BrdU-positive nuclei were distributed mainly in the “pinwheel” architecture of the ventricular epithelium (A). The DCX+ cells occupied an adjacent, distinct position [(A), arrows]. Some of the DCX+ migrated away from the ventricular wall (B). We noted vigorous neurogenesis with many DCX+ (arrows) co-localizing with BrdU nuclei in the G-CSF-treated animals [(C); arrowheads] and animals treated with G-CSF + BM MSC [(D), arrows]. (E–G) Post-stroke angiogenesis. In regions adjacent to the infarct scar, we found numerous BrdU+ nuclei in the endothelium of newly formed blood vessels in the formerly infarct core [(E), green]. The border to the healthy brain region was abruptly demarcated to the left by NeuN-positive nuclei [(E), red]. Beyond the formerly infarct core, we noted vigorous sprouting angiogenesis as evidenced by RECA/BrdU double positive blood vessels [(F), violet] as well as numerous BrdU+ nuclei in the newly formed endothelium [(F), blue] and reconstruction of the basal lamina [(F), green] during the resolution phase of angiogenesis. By number of laminin/BrdU co-localizations, the density of the newly formed blood vessels was significantly higher (threefold, p = 0.01) in the brains of animals treated with the combination G-CSF + BM MSC as compared to controls and G-CSF alone (G). Cc, corpus callosum; IC, infarct core; IR, islet of regeneration; LV, lateral ventricle; PI, periinfract.

Mentions: Next, we investigated the presence of the early neuronal marker doublecortin by immunofluorescence in the lateral ventricle region. To this end, the proliferating cells were labeled by injecting animals with BrdU. To our surprise, at day 56 post-stroke none of the DCX+ cells in the SVZ of control animals co-localized with BrdU-labeled nuclei. Instead, the BrdU-positive nuclei were distributed mainly in the “pinwheel” architecture of the ventricular epithelium (Liebner et al., 2008; Gajera et al., 2010) (Figure 5A). The DCX+ cells occupied an adjacent, distinct position (Figure 5A, arrows). Some of the DCX+ migrated away from the ventricular wall (Figure 5B). In agreement with the previous results, we noted vigorous neurogenesis with many DCX+ (arrows) co-localizing with BrdU nuclei in the G-CSF-treated animals (Figure 5C; arrowheads) and animals treated with G-CSF + BM MSC (Figure 5D, arrows).


Multimodal Approaches for Regenerative Stroke Therapies: Combination of Granulocyte Colony-Stimulating Factor with Bone Marrow Mesenchymal Stem Cells is Not Superior to G-CSF Alone.

Balseanu AT, Buga AM, Catalin B, Wagner DC, Boltze J, Zagrean AM, Reymann K, Schaebitz W, Popa-Wagner A - Front Aging Neurosci (2014)

Post-stroke neurogenesis and angiogenesis. At 8 weeks post-stroke, none of the DCX+ cells in the SVZ of control animals co-localized with BrdU-labeled nuclei. Instead, the BrdU-positive nuclei were distributed mainly in the “pinwheel” architecture of the ventricular epithelium (A). The DCX+ cells occupied an adjacent, distinct position [(A), arrows]. Some of the DCX+ migrated away from the ventricular wall (B). We noted vigorous neurogenesis with many DCX+ (arrows) co-localizing with BrdU nuclei in the G-CSF-treated animals [(C); arrowheads] and animals treated with G-CSF + BM MSC [(D), arrows]. (E–G) Post-stroke angiogenesis. In regions adjacent to the infarct scar, we found numerous BrdU+ nuclei in the endothelium of newly formed blood vessels in the formerly infarct core [(E), green]. The border to the healthy brain region was abruptly demarcated to the left by NeuN-positive nuclei [(E), red]. Beyond the formerly infarct core, we noted vigorous sprouting angiogenesis as evidenced by RECA/BrdU double positive blood vessels [(F), violet] as well as numerous BrdU+ nuclei in the newly formed endothelium [(F), blue] and reconstruction of the basal lamina [(F), green] during the resolution phase of angiogenesis. By number of laminin/BrdU co-localizations, the density of the newly formed blood vessels was significantly higher (threefold, p = 0.01) in the brains of animals treated with the combination G-CSF + BM MSC as compared to controls and G-CSF alone (G). Cc, corpus callosum; IC, infarct core; IR, islet of regeneration; LV, lateral ventricle; PI, periinfract.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 5: Post-stroke neurogenesis and angiogenesis. At 8 weeks post-stroke, none of the DCX+ cells in the SVZ of control animals co-localized with BrdU-labeled nuclei. Instead, the BrdU-positive nuclei were distributed mainly in the “pinwheel” architecture of the ventricular epithelium (A). The DCX+ cells occupied an adjacent, distinct position [(A), arrows]. Some of the DCX+ migrated away from the ventricular wall (B). We noted vigorous neurogenesis with many DCX+ (arrows) co-localizing with BrdU nuclei in the G-CSF-treated animals [(C); arrowheads] and animals treated with G-CSF + BM MSC [(D), arrows]. (E–G) Post-stroke angiogenesis. In regions adjacent to the infarct scar, we found numerous BrdU+ nuclei in the endothelium of newly formed blood vessels in the formerly infarct core [(E), green]. The border to the healthy brain region was abruptly demarcated to the left by NeuN-positive nuclei [(E), red]. Beyond the formerly infarct core, we noted vigorous sprouting angiogenesis as evidenced by RECA/BrdU double positive blood vessels [(F), violet] as well as numerous BrdU+ nuclei in the newly formed endothelium [(F), blue] and reconstruction of the basal lamina [(F), green] during the resolution phase of angiogenesis. By number of laminin/BrdU co-localizations, the density of the newly formed blood vessels was significantly higher (threefold, p = 0.01) in the brains of animals treated with the combination G-CSF + BM MSC as compared to controls and G-CSF alone (G). Cc, corpus callosum; IC, infarct core; IR, islet of regeneration; LV, lateral ventricle; PI, periinfract.
Mentions: Next, we investigated the presence of the early neuronal marker doublecortin by immunofluorescence in the lateral ventricle region. To this end, the proliferating cells were labeled by injecting animals with BrdU. To our surprise, at day 56 post-stroke none of the DCX+ cells in the SVZ of control animals co-localized with BrdU-labeled nuclei. Instead, the BrdU-positive nuclei were distributed mainly in the “pinwheel” architecture of the ventricular epithelium (Liebner et al., 2008; Gajera et al., 2010) (Figure 5A). The DCX+ cells occupied an adjacent, distinct position (Figure 5A, arrows). Some of the DCX+ migrated away from the ventricular wall (Figure 5B). In agreement with the previous results, we noted vigorous neurogenesis with many DCX+ (arrows) co-localizing with BrdU nuclei in the G-CSF-treated animals (Figure 5C; arrowheads) and animals treated with G-CSF + BM MSC (Figure 5D, arrows).

Bottom Line: Functional recovery was tested during the entire post-stroke survival period of 56 days.The combination therapy also led to robust angiogenesis in the formerly infarct core and beyond in the "islet of regeneration." However, G-CSF + BM MSCs may not impact at all on the spatial reference-memory task or infarct volume and therefore did not further improve the post-stroke recovery.We suggest that in a real clinical practice involving older post-stroke patients, successful regenerative therapies would have to be carried out for a much longer time.

View Article: PubMed Central - PubMed

Affiliation: Center of Clinical and Experimental Medicine, University of Medicine and Pharmacy of Craiova , Craiova , Romania.

ABSTRACT
Attractive therapeutic strategies to enhance post-stroke recovery of aged brains include methods of cellular therapy that can enhance the endogenous restorative mechanisms of the injured brain. Since stroke afflicts mostly the elderly, it is highly desirable to test the efficacy of cell therapy in the microenvironment of aged brains that is generally refractory to regeneration. In particular, stem cells from the bone marrow allow an autologous transplantation approach that can be translated in the near future to the clinical practice. Such a bone marrow-derived therapy includes the grafting of stem cells as well as the delayed induction of endogenous stem cell mobilization and homing by the stem cell mobilizer granulocyte colony-stimulating factor (G-CSF). We tested the hypothesis that grafting of bone marrow-derived pre-differentiated mesenchymal cells (BM-MSCs) in G-CSF-treated animals improves the long-term functional outcome in aged rodents. To this end, G-CSF alone (50 μg/kg) or in combination with a single dose (10(6) cells) of rat BM MSCs was administered intravenously to Sprague-Dawley rats at 6 h after transient occlusion (90 min) of the middle cerebral artery. Infarct volume was measured by magnetic resonance imaging at 3 and 48 days post-stroke and additionally by immunhistochemistry at day 56. Functional recovery was tested during the entire post-stroke survival period of 56 days. Daily treatment for post-stroke aged rats with G-CSF led to a robust and consistent improvement of neurological function after 28 days. The combination therapy also led to robust angiogenesis in the formerly infarct core and beyond in the "islet of regeneration." However, G-CSF + BM MSCs may not impact at all on the spatial reference-memory task or infarct volume and therefore did not further improve the post-stroke recovery. We suggest that in a real clinical practice involving older post-stroke patients, successful regenerative therapies would have to be carried out for a much longer time.

No MeSH data available.


Related in: MedlinePlus