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Transplanted Neural Progenitor Cells from Distinct Sources Migrate Differentially in an Organotypic Model of Brain Injury.

Ngalula KP, Cramer N, Schell MJ, Juliano SL - Front Neurol (2015)

Bottom Line: NPCs derived from the GE tended to be immunoreactive for GABAergic markers while those derived from the neocortex were more strongly immunoreactive for other neuronal markers such as MAP2, TUJ1, or Milli-Mark.Cells transplanted in vitro acquired the electrophysiological characteristics of neurons, including action potential generation and reception of spontaneous synaptic activity.This suggests that transplanted cells differentiate into neurons capable of functionally integrating with the host tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Physiology and Genetics, Uniformed Services University of Health Sciences , Bethesda, MD , USA.

ABSTRACT
Brain injury is a major cause of long-term disability. The possibility exists for exogenously derived neural progenitor cells to repair damage resulting from brain injury, although more information is needed to successfully implement this promising therapy. To test the ability of neural progenitor cells (NPCs) obtained from rats to repair damaged neocortex, we transplanted neural progenitor cell suspensions into normal and injured slice cultures of the neocortex acquired from rats on postnatal day 0-3. Donor cells from E16 embryos were obtained from either the neocortex, including the ventricular zone (VZ) for excitatory cells, ganglionic eminence (GE) for inhibitory cells or a mixed population of the two. Cells were injected into the ventricular/subventricular zone (VZ/SVZ) or directly into the wounded region. Transplanted cells migrated throughout the cortical plate with GE and mixed population donor cells predominately targeting the upper cortical layers, while neocortically derived NPCs from the VZ/SVZ migrated less extensively. In the injured neocortex, transplanted cells moved predominantly into the wounded area. NPCs derived from the GE tended to be immunoreactive for GABAergic markers while those derived from the neocortex were more strongly immunoreactive for other neuronal markers such as MAP2, TUJ1, or Milli-Mark. Cells transplanted in vitro acquired the electrophysiological characteristics of neurons, including action potential generation and reception of spontaneous synaptic activity. This suggests that transplanted cells differentiate into neurons capable of functionally integrating with the host tissue. Together, our data suggest that transplantation of neural progenitor cells holds great potential as an emerging therapeutic intervention for restoring function lost to brain damage.

No MeSH data available.


Related in: MedlinePlus

Transplanted cells immunoreactive for different markers. This graph represents the percentage of the total number of transplanted cells labeled with CMDiI that were counted for each marker, which were also double labeled for a specific antibody. The total number of counted cells for each marker can be seen in Table 2. In general, similar numbers of transplanted cells from derived from each source (GE, Ctx, Mixed) were immunoreactive for the neuronal markers, Tuj1 (TUJ) and Milli-Mark. GE-derived cells were more likely to differentiate into cells immunoreactive for GABA, while neocortically derived cells are more immunoreactive for MAP2 (*p < 0.01). Very few transplanted cells from any source are immunoreactive for GFAP (#p > 0.05). We used a two-way ANOVA followed by pairwise comparisons with the Holm–Sidak test, error bars = SEM.
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Figure 6: Transplanted cells immunoreactive for different markers. This graph represents the percentage of the total number of transplanted cells labeled with CMDiI that were counted for each marker, which were also double labeled for a specific antibody. The total number of counted cells for each marker can be seen in Table 2. In general, similar numbers of transplanted cells from derived from each source (GE, Ctx, Mixed) were immunoreactive for the neuronal markers, Tuj1 (TUJ) and Milli-Mark. GE-derived cells were more likely to differentiate into cells immunoreactive for GABA, while neocortically derived cells are more immunoreactive for MAP2 (*p < 0.01). Very few transplanted cells from any source are immunoreactive for GFAP (#p > 0.05). We used a two-way ANOVA followed by pairwise comparisons with the Holm–Sidak test, error bars = SEM.

Mentions: To further characterize the phenotype of the transplanted cells, the organotypic culture slices were fixed at day 5 or 7 post transplantation and immunoreacted for neuronal and glial markers. The distinct morphologies of the transplanted cells, as shown in Figure 2D, suggests that they were differentiating into well-defined neural cell types. To more completely assign a phenotype to the transplanted cells, we used a battery of markers to further characterize their identity. In addition, the NPCs transplanted from different embryonic sources might differentiate into distinct neuronal types. We used several neural markers: GABA, MAP2, β-tubulin III (TUJ-1), Milli-Mark, and GFAP (Figure 5). The labeled transplanted cells showed immunoreactivity for multiple markers, suggesting they differentiated into a variety of cell types. Figure 5 demonstrates transplanted cells labeled with CMDiI or GFP and immunoreactive for neuronal (Figures 5A–L,P–R) or glial markers (Figures 5M–O). Figures 5A–C shows a cell labeled with CMDiI displaying a migratory morphology immunoreactive against GABA. The images in Figures 5D–L illustrate examples of labeled transplanted cells reactive for other neuronal markers, including MAP2 (D–F and G–I), Milli-Mark (J–L) and TUJ (P–R). Figures 5M–O shows an example of a CMDiI labeled cell surrounded by GFAP immunoreactive cells, demonstrating that many GFAP+ cells occurred in each organotypic slice, but very few of the transplanted cells were GFAP+. Figure 6 shows the percent of transplanted cells immunoreactive for all markers across experiments and also illustrates the fraction of cells from each source that displayed immunoreactivity out of the total number of cells that were transplanted. Transplants derived from the GE were more likely to differentiate into GABAergic cells than neocortically derived cells. All populations of cells demonstrated very low reactivity for GFAP, suggesting that very few of the transplanted cells differentiate into astrocytes. The numbers of cells counted and used to produce Figure 6 can be seen in Table 3.


Transplanted Neural Progenitor Cells from Distinct Sources Migrate Differentially in an Organotypic Model of Brain Injury.

Ngalula KP, Cramer N, Schell MJ, Juliano SL - Front Neurol (2015)

Transplanted cells immunoreactive for different markers. This graph represents the percentage of the total number of transplanted cells labeled with CMDiI that were counted for each marker, which were also double labeled for a specific antibody. The total number of counted cells for each marker can be seen in Table 2. In general, similar numbers of transplanted cells from derived from each source (GE, Ctx, Mixed) were immunoreactive for the neuronal markers, Tuj1 (TUJ) and Milli-Mark. GE-derived cells were more likely to differentiate into cells immunoreactive for GABA, while neocortically derived cells are more immunoreactive for MAP2 (*p < 0.01). Very few transplanted cells from any source are immunoreactive for GFAP (#p > 0.05). We used a two-way ANOVA followed by pairwise comparisons with the Holm–Sidak test, error bars = SEM.
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Figure 6: Transplanted cells immunoreactive for different markers. This graph represents the percentage of the total number of transplanted cells labeled with CMDiI that were counted for each marker, which were also double labeled for a specific antibody. The total number of counted cells for each marker can be seen in Table 2. In general, similar numbers of transplanted cells from derived from each source (GE, Ctx, Mixed) were immunoreactive for the neuronal markers, Tuj1 (TUJ) and Milli-Mark. GE-derived cells were more likely to differentiate into cells immunoreactive for GABA, while neocortically derived cells are more immunoreactive for MAP2 (*p < 0.01). Very few transplanted cells from any source are immunoreactive for GFAP (#p > 0.05). We used a two-way ANOVA followed by pairwise comparisons with the Holm–Sidak test, error bars = SEM.
Mentions: To further characterize the phenotype of the transplanted cells, the organotypic culture slices were fixed at day 5 or 7 post transplantation and immunoreacted for neuronal and glial markers. The distinct morphologies of the transplanted cells, as shown in Figure 2D, suggests that they were differentiating into well-defined neural cell types. To more completely assign a phenotype to the transplanted cells, we used a battery of markers to further characterize their identity. In addition, the NPCs transplanted from different embryonic sources might differentiate into distinct neuronal types. We used several neural markers: GABA, MAP2, β-tubulin III (TUJ-1), Milli-Mark, and GFAP (Figure 5). The labeled transplanted cells showed immunoreactivity for multiple markers, suggesting they differentiated into a variety of cell types. Figure 5 demonstrates transplanted cells labeled with CMDiI or GFP and immunoreactive for neuronal (Figures 5A–L,P–R) or glial markers (Figures 5M–O). Figures 5A–C shows a cell labeled with CMDiI displaying a migratory morphology immunoreactive against GABA. The images in Figures 5D–L illustrate examples of labeled transplanted cells reactive for other neuronal markers, including MAP2 (D–F and G–I), Milli-Mark (J–L) and TUJ (P–R). Figures 5M–O shows an example of a CMDiI labeled cell surrounded by GFAP immunoreactive cells, demonstrating that many GFAP+ cells occurred in each organotypic slice, but very few of the transplanted cells were GFAP+. Figure 6 shows the percent of transplanted cells immunoreactive for all markers across experiments and also illustrates the fraction of cells from each source that displayed immunoreactivity out of the total number of cells that were transplanted. Transplants derived from the GE were more likely to differentiate into GABAergic cells than neocortically derived cells. All populations of cells demonstrated very low reactivity for GFAP, suggesting that very few of the transplanted cells differentiate into astrocytes. The numbers of cells counted and used to produce Figure 6 can be seen in Table 3.

Bottom Line: NPCs derived from the GE tended to be immunoreactive for GABAergic markers while those derived from the neocortex were more strongly immunoreactive for other neuronal markers such as MAP2, TUJ1, or Milli-Mark.Cells transplanted in vitro acquired the electrophysiological characteristics of neurons, including action potential generation and reception of spontaneous synaptic activity.This suggests that transplanted cells differentiate into neurons capable of functionally integrating with the host tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Physiology and Genetics, Uniformed Services University of Health Sciences , Bethesda, MD , USA.

ABSTRACT
Brain injury is a major cause of long-term disability. The possibility exists for exogenously derived neural progenitor cells to repair damage resulting from brain injury, although more information is needed to successfully implement this promising therapy. To test the ability of neural progenitor cells (NPCs) obtained from rats to repair damaged neocortex, we transplanted neural progenitor cell suspensions into normal and injured slice cultures of the neocortex acquired from rats on postnatal day 0-3. Donor cells from E16 embryos were obtained from either the neocortex, including the ventricular zone (VZ) for excitatory cells, ganglionic eminence (GE) for inhibitory cells or a mixed population of the two. Cells were injected into the ventricular/subventricular zone (VZ/SVZ) or directly into the wounded region. Transplanted cells migrated throughout the cortical plate with GE and mixed population donor cells predominately targeting the upper cortical layers, while neocortically derived NPCs from the VZ/SVZ migrated less extensively. In the injured neocortex, transplanted cells moved predominantly into the wounded area. NPCs derived from the GE tended to be immunoreactive for GABAergic markers while those derived from the neocortex were more strongly immunoreactive for other neuronal markers such as MAP2, TUJ1, or Milli-Mark. Cells transplanted in vitro acquired the electrophysiological characteristics of neurons, including action potential generation and reception of spontaneous synaptic activity. This suggests that transplanted cells differentiate into neurons capable of functionally integrating with the host tissue. Together, our data suggest that transplantation of neural progenitor cells holds great potential as an emerging therapeutic intervention for restoring function lost to brain damage.

No MeSH data available.


Related in: MedlinePlus