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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

The distribution of transplanted cells in normal cultures. (A) shows the mediolateral distribution of cells transplanted into normal cortex after 7 days in culture. Significantly fewer cells reached lateral portions of the cortical slice compared with transplanted cells migrating into the middle and medial regions of the slice. (B) illustrates the distribution of cells that migrated away from the injection site. The bars to the right of the vertical line show that of the cells migrating away from the injection site, a greater percentage reached the cortical plate (CP) than those remaining in the intermediate zone (IZ). Significantly fewer cells obtained from the neocortex alone, however, reached the cortical plate compared with cells obtained from the GE or the mixed population (*p < 0.01). The bars to the left of the vertical line show the distribution of cells that reached the cortical plate and moved into the upper (U), middle (M), or lower (L) cortical layers. Of the cells that reach the cortical plate, fewer cells resided in the lower layers of the cortical plate (#p < 0.05). Compared with the GE and mixed populations, a significantly smaller percentage of neocortically derived cells reached the upper layers (*p < 0.001) and a greater percentage of cells remained in the lower layers (*p < 0.001). See Table 1 for the numbers of cells in each group. (Two-way ANOVA followed by the Holm–Sidak pairwise comparison, error bars = SEM).
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Figure 3: The distribution of transplanted cells in normal cultures. (A) shows the mediolateral distribution of cells transplanted into normal cortex after 7 days in culture. Significantly fewer cells reached lateral portions of the cortical slice compared with transplanted cells migrating into the middle and medial regions of the slice. (B) illustrates the distribution of cells that migrated away from the injection site. The bars to the right of the vertical line show that of the cells migrating away from the injection site, a greater percentage reached the cortical plate (CP) than those remaining in the intermediate zone (IZ). Significantly fewer cells obtained from the neocortex alone, however, reached the cortical plate compared with cells obtained from the GE or the mixed population (*p < 0.01). The bars to the left of the vertical line show the distribution of cells that reached the cortical plate and moved into the upper (U), middle (M), or lower (L) cortical layers. Of the cells that reach the cortical plate, fewer cells resided in the lower layers of the cortical plate (#p < 0.05). Compared with the GE and mixed populations, a significantly smaller percentage of neocortically derived cells reached the upper layers (*p < 0.001) and a greater percentage of cells remained in the lower layers (*p < 0.001). See Table 1 for the numbers of cells in each group. (Two-way ANOVA followed by the Holm–Sidak pairwise comparison, error bars = SEM).

Mentions: After the slices were placed in culture, they received transplants of acutely derived cell suspensions obtained from the neocortex alone, the GE alone, or a mixed population of neocortex and GE. Based on the count of viable transplanted cells and the number of transplanted cells, we approximated the number of cells that migrated away from the injection site. About 20% of the total amount of transplanted cells migrated away from an injection suggesting a similar viability percentage. In normal uninjured slices, each cell population moved into the cortical plate and migrated extensively into medial, middle, and lateral parts of the slices (Figures 2A–C; also see Table 1). After reaching their destination, the transplanted cells acquired different morphologies and displayed multiple processes and neuron-like morphologies (Figure 2D). All of the transplanted cell types (neocortical, GE, and mixed) distributed similarly in the different regions of the host slice. When we quantified the medial to lateral distribution of transplanted cells, we observed that cells of all groups migrated in greater numbers toward the medial portions of the host cortical slice, whereas fewer cells moved into lateral regions (Figure 3A; Table 1). More cells derived from any source moved into the middle and medial regions of the CP (Figure 3A). Most of the migrating cells moved through the intermediate zone into the cortical plate (Figure 3B). More cells derived from the GE and mixed cell population, however, reached the cortical plate than those derived from the neocortex, leaving a greater percentage of neocortically derived cells in the intermediate zone (Figure 3B; Table 1). In the CP, GE cells are in great number located into the upper layer while neocortical cells are preferentially in the middle and lower layers. These results suggest that: (i) all populations of transplanted cells (GE-derived, neocortical-derived, and mixed) migrate preferentially toward the middle and medial CP, (ii) GE-derived cells migrate more efficiently than cortical cells, and (iii) the migration of ­neocortically derived cells is improved when transplanted together with GE-derived cells.


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)

The distribution of transplanted cells in normal cultures. (A) shows the mediolateral distribution of cells transplanted into normal cortex after 7 days in culture. Significantly fewer cells reached lateral portions of the cortical slice compared with transplanted cells migrating into the middle and medial regions of the slice. (B) illustrates the distribution of cells that migrated away from the injection site. The bars to the right of the vertical line show that of the cells migrating away from the injection site, a greater percentage reached the cortical plate (CP) than those remaining in the intermediate zone (IZ). Significantly fewer cells obtained from the neocortex alone, however, reached the cortical plate compared with cells obtained from the GE or the mixed population (*p < 0.01). The bars to the left of the vertical line show the distribution of cells that reached the cortical plate and moved into the upper (U), middle (M), or lower (L) cortical layers. Of the cells that reach the cortical plate, fewer cells resided in the lower layers of the cortical plate (#p < 0.05). Compared with the GE and mixed populations, a significantly smaller percentage of neocortically derived cells reached the upper layers (*p < 0.001) and a greater percentage of cells remained in the lower layers (*p < 0.001). See Table 1 for the numbers of cells in each group. (Two-way ANOVA followed by the Holm–Sidak pairwise comparison, error bars = SEM).
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Figure 3: The distribution of transplanted cells in normal cultures. (A) shows the mediolateral distribution of cells transplanted into normal cortex after 7 days in culture. Significantly fewer cells reached lateral portions of the cortical slice compared with transplanted cells migrating into the middle and medial regions of the slice. (B) illustrates the distribution of cells that migrated away from the injection site. The bars to the right of the vertical line show that of the cells migrating away from the injection site, a greater percentage reached the cortical plate (CP) than those remaining in the intermediate zone (IZ). Significantly fewer cells obtained from the neocortex alone, however, reached the cortical plate compared with cells obtained from the GE or the mixed population (*p < 0.01). The bars to the left of the vertical line show the distribution of cells that reached the cortical plate and moved into the upper (U), middle (M), or lower (L) cortical layers. Of the cells that reach the cortical plate, fewer cells resided in the lower layers of the cortical plate (#p < 0.05). Compared with the GE and mixed populations, a significantly smaller percentage of neocortically derived cells reached the upper layers (*p < 0.001) and a greater percentage of cells remained in the lower layers (*p < 0.001). See Table 1 for the numbers of cells in each group. (Two-way ANOVA followed by the Holm–Sidak pairwise comparison, error bars = SEM).
Mentions: After the slices were placed in culture, they received transplants of acutely derived cell suspensions obtained from the neocortex alone, the GE alone, or a mixed population of neocortex and GE. Based on the count of viable transplanted cells and the number of transplanted cells, we approximated the number of cells that migrated away from the injection site. About 20% of the total amount of transplanted cells migrated away from an injection suggesting a similar viability percentage. In normal uninjured slices, each cell population moved into the cortical plate and migrated extensively into medial, middle, and lateral parts of the slices (Figures 2A–C; also see Table 1). After reaching their destination, the transplanted cells acquired different morphologies and displayed multiple processes and neuron-like morphologies (Figure 2D). All of the transplanted cell types (neocortical, GE, and mixed) distributed similarly in the different regions of the host slice. When we quantified the medial to lateral distribution of transplanted cells, we observed that cells of all groups migrated in greater numbers toward the medial portions of the host cortical slice, whereas fewer cells moved into lateral regions (Figure 3A; Table 1). More cells derived from any source moved into the middle and medial regions of the CP (Figure 3A). Most of the migrating cells moved through the intermediate zone into the cortical plate (Figure 3B). More cells derived from the GE and mixed cell population, however, reached the cortical plate than those derived from the neocortex, leaving a greater percentage of neocortically derived cells in the intermediate zone (Figure 3B; Table 1). In the CP, GE cells are in great number located into the upper layer while neocortical cells are preferentially in the middle and lower layers. These results suggest that: (i) all populations of transplanted cells (GE-derived, neocortical-derived, and mixed) migrate preferentially toward the middle and medial CP, (ii) GE-derived cells migrate more efficiently than cortical cells, and (iii) the migration of ­neocortically derived cells is improved when transplanted together with GE-derived cells.

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