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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

Model of injury and transplantation. (A) is an organotypic culture obtained on postnatal day 1 (P1), that also sustained an injury. The arrow is the approximate site of a transplant into the SVZ/VZ. (A′) shows the boxed area in (A) and contains a higher power view of the injury outlined by arrowheads; the arrow points to the deep end of the lesion. (B) is an organotypic culture obtained at P1 without injury. For analysis, the neocortex was divided into regions designated as medial, middle, and lateral. (B′) is a higher magnification of the boxed in region in (B) and delineates laminar distinctions in the cortical plate used to define the positions of the transplanted migrating cells. The arrow in (A′) represents the approximate site of cells injected into a lesion and the arrow head in (B′) represents the approximate site of cells injected into the VZ/SVZ of a slice with no lesion. (C) is a coronal section of E16 brain, (C′) is higher magnification of the boxed in region in (C) and represents the developing cortical wall. This is the region used for preparing the cell suspension made from the embryonic neocortex. The circle in C encloses the site used for preparing the GE cell suspension. CP: cortical plate, U: upper layer, M: middle layer, L: lower layer, IZ: intermediate zone, SVZ, subventricular zone; VZ, ventricular zone; LV, lateral ventricle. Scale bar = 500 μm (A–C), 100 μm (A′–C′).
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Figure 1: Model of injury and transplantation. (A) is an organotypic culture obtained on postnatal day 1 (P1), that also sustained an injury. The arrow is the approximate site of a transplant into the SVZ/VZ. (A′) shows the boxed area in (A) and contains a higher power view of the injury outlined by arrowheads; the arrow points to the deep end of the lesion. (B) is an organotypic culture obtained at P1 without injury. For analysis, the neocortex was divided into regions designated as medial, middle, and lateral. (B′) is a higher magnification of the boxed in region in (B) and delineates laminar distinctions in the cortical plate used to define the positions of the transplanted migrating cells. The arrow in (A′) represents the approximate site of cells injected into a lesion and the arrow head in (B′) represents the approximate site of cells injected into the VZ/SVZ of a slice with no lesion. (C) is a coronal section of E16 brain, (C′) is higher magnification of the boxed in region in (C) and represents the developing cortical wall. This is the region used for preparing the cell suspension made from the embryonic neocortex. The circle in C encloses the site used for preparing the GE cell suspension. CP: cortical plate, U: upper layer, M: middle layer, L: lower layer, IZ: intermediate zone, SVZ, subventricular zone; VZ, ventricular zone; LV, lateral ventricle. Scale bar = 500 μm (A–C), 100 μm (A′–C′).

Mentions: An example of an organotypic culture containing a lesion can be seen in Figure 1A. For analysis, the slice was divided into medial, middle, and lateral regions (Figure 1B). The immature cortical plate was divided into upper, middle, and lower layers (Figure 1B′). The approximate sites of transplantation are indicated by an arrow (Figures 1A,A′,B′). Cells obtained acutely from E16 rat embryos (Figures 1C,C′) were transplanted into the VZ/SVZ of a neocortical slice in control and injured cultures as indicated by the arrows in Figures 1A–B′.


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)

Model of injury and transplantation. (A) is an organotypic culture obtained on postnatal day 1 (P1), that also sustained an injury. The arrow is the approximate site of a transplant into the SVZ/VZ. (A′) shows the boxed area in (A) and contains a higher power view of the injury outlined by arrowheads; the arrow points to the deep end of the lesion. (B) is an organotypic culture obtained at P1 without injury. For analysis, the neocortex was divided into regions designated as medial, middle, and lateral. (B′) is a higher magnification of the boxed in region in (B) and delineates laminar distinctions in the cortical plate used to define the positions of the transplanted migrating cells. The arrow in (A′) represents the approximate site of cells injected into a lesion and the arrow head in (B′) represents the approximate site of cells injected into the VZ/SVZ of a slice with no lesion. (C) is a coronal section of E16 brain, (C′) is higher magnification of the boxed in region in (C) and represents the developing cortical wall. This is the region used for preparing the cell suspension made from the embryonic neocortex. The circle in C encloses the site used for preparing the GE cell suspension. CP: cortical plate, U: upper layer, M: middle layer, L: lower layer, IZ: intermediate zone, SVZ, subventricular zone; VZ, ventricular zone; LV, lateral ventricle. Scale bar = 500 μm (A–C), 100 μm (A′–C′).
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Related In: Results  -  Collection

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Figure 1: Model of injury and transplantation. (A) is an organotypic culture obtained on postnatal day 1 (P1), that also sustained an injury. The arrow is the approximate site of a transplant into the SVZ/VZ. (A′) shows the boxed area in (A) and contains a higher power view of the injury outlined by arrowheads; the arrow points to the deep end of the lesion. (B) is an organotypic culture obtained at P1 without injury. For analysis, the neocortex was divided into regions designated as medial, middle, and lateral. (B′) is a higher magnification of the boxed in region in (B) and delineates laminar distinctions in the cortical plate used to define the positions of the transplanted migrating cells. The arrow in (A′) represents the approximate site of cells injected into a lesion and the arrow head in (B′) represents the approximate site of cells injected into the VZ/SVZ of a slice with no lesion. (C) is a coronal section of E16 brain, (C′) is higher magnification of the boxed in region in (C) and represents the developing cortical wall. This is the region used for preparing the cell suspension made from the embryonic neocortex. The circle in C encloses the site used for preparing the GE cell suspension. CP: cortical plate, U: upper layer, M: middle layer, L: lower layer, IZ: intermediate zone, SVZ, subventricular zone; VZ, ventricular zone; LV, lateral ventricle. Scale bar = 500 μm (A–C), 100 μm (A′–C′).
Mentions: An example of an organotypic culture containing a lesion can be seen in Figure 1A. For analysis, the slice was divided into medial, middle, and lateral regions (Figure 1B). The immature cortical plate was divided into upper, middle, and lower layers (Figure 1B′). The approximate sites of transplantation are indicated by an arrow (Figures 1A,A′,B′). Cells obtained acutely from E16 rat embryos (Figures 1C,C′) were transplanted into the VZ/SVZ of a neocortical slice in control and injured cultures as indicated by the arrows in Figures 1A–B′.

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