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Human fetal brain-derived neural stem/progenitor cells grafted into the adult epileptic brain restrain seizures in rat models of temporal lobe epilepsy.

Lee H, Yun S, Kim IS, Lee IS, Shin JE, Park SC, Kim WJ, Park KI - PLoS ONE (2014)

Bottom Line: However, NSPC grafting neither improved spatial learning or memory function in pilocarpine-treated animals.Grafted cells restored the expression of GDNF in host astrocytes but did not reverse the mossy fiber sprouting, eliminating the latter as potential mechanism.These results suggest that human fetal brain-derived NSPCs possess some therapeutic effect for TLE treatments although further studies to both increase the yield of NSPC grafts-derived functionally integrated GABAergic neurons and improve cognitive deficits are still needed.

View Article: PubMed Central - PubMed

Affiliation: Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT
Cell transplantation has been suggested as an alternative therapy for temporal lobe epilepsy (TLE) because this can suppress spontaneous recurrent seizures in animal models. To evaluate the therapeutic potential of human neural stem/progenitor cells (huNSPCs) for treating TLE, we transplanted huNSPCs, derived from an aborted fetal telencephalon at 13 weeks of gestation and expanded in culture as neurospheres over a long time period, into the epileptic hippocampus of fully kindled and pilocarpine-treated adult rats exhibiting TLE. In vitro, huNSPCs not only produced all three central nervous system neural cell types, but also differentiated into ganglionic eminences-derived γ-aminobutyric acid (GABA)-ergic interneurons and released GABA in response to the depolarization induced by a high K+ medium. NSPC grafting reduced behavioral seizure duration, afterdischarge duration on electroencephalograms, and seizure stage in the kindling model, as well as the frequency and the duration of spontaneous recurrent motor seizures in pilocarpine-induced animals. However, NSPC grafting neither improved spatial learning or memory function in pilocarpine-treated animals. Following transplantation, grafted cells showed extensive migration around the injection site, robust engraftment, and long-term survival, along with differentiation into β-tubulin III+ neurons (∼34%), APC-CC1+ oligodendrocytes (∼28%), and GFAP+ astrocytes (∼8%). Furthermore, among donor-derived cells, ∼24% produced GABA. Additionally, to explain the effect of seizure suppression after NSPC grafting, we examined the anticonvulsant glial cell-derived neurotrophic factor (GDNF) levels in host hippocampal astrocytes and mossy fiber sprouting into the supragranular layer of the dentate gyrus in the epileptic brain. Grafted cells restored the expression of GDNF in host astrocytes but did not reverse the mossy fiber sprouting, eliminating the latter as potential mechanism. These results suggest that human fetal brain-derived NSPCs possess some therapeutic effect for TLE treatments although further studies to both increase the yield of NSPC grafts-derived functionally integrated GABAergic neurons and improve cognitive deficits are still needed.

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Differentiation of human NSPCs following transplantation into the hippocampus of kindled rats.(A–F) BrdU+ grafted cells (green) were co-stained with TUJ1 (red, arrowheads in B, C, E, F) in the CA3 region of the hippocampus (A–C) and fimbria (D–F) in rats. (G–J) hNP+ grafted cells (red, arrowheads in G, I, J) were co-localized with TUJ1 (green, arrowheads in H–J) in the hilus of the hippocampus. (J) Orthogonal view from confocal z-series showed that hNP (red) in nuclei and TUJ1 (green) in cytoplasm were expressed in the same cell. (K, M) Under the dual-filter microscope, BrdU+ grafted cells co-expressed APC-CC1 in the fimbria (arrows in K), and hNuc+ grafted cells co-expressed GFAP in the CA3 region (an arrow in M). Scale bar; 200 µm (A), 20 µm (D), 10 µm (G, J), 20 µm (K).
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pone-0104092-g005: Differentiation of human NSPCs following transplantation into the hippocampus of kindled rats.(A–F) BrdU+ grafted cells (green) were co-stained with TUJ1 (red, arrowheads in B, C, E, F) in the CA3 region of the hippocampus (A–C) and fimbria (D–F) in rats. (G–J) hNP+ grafted cells (red, arrowheads in G, I, J) were co-localized with TUJ1 (green, arrowheads in H–J) in the hilus of the hippocampus. (J) Orthogonal view from confocal z-series showed that hNP (red) in nuclei and TUJ1 (green) in cytoplasm were expressed in the same cell. (K, M) Under the dual-filter microscope, BrdU+ grafted cells co-expressed APC-CC1 in the fimbria (arrows in K), and hNuc+ grafted cells co-expressed GFAP in the CA3 region (an arrow in M). Scale bar; 200 µm (A), 20 µm (D), 10 µm (G, J), 20 µm (K).

Mentions: To study differentiation patterns of donor-derived cells, the whole hippocampal area of kindled rats was analyzed at 8 weeks following transplantation. BrdU+ cells were co-labeled with the early neuronal marker TUJ1 in the CA3 region of the hippocampus (Fig. 5A–C) and fimbria (Fig. 5D–F). Confocal microscopic images of hNP+-grafted cells demonstrated dual labeling with the TUJ1 antibody (Fig. 5G–J). Quantification revealed that donor-derived cells differentiated into TUJ1+ neurons (33.8±4.0%, n = 4), APC-CC1-expressing oligodendrocytes (27.7±6.2%, n = 4), and GFAP-expressing astrocytes (8.2±1.8%, n = 4) in the hippocampus (Fig. 5K, M). About 40% of GFAP-expressing astrocytes expressed GDNF (Fig. S3). Additionally, ∼24% of BrdU+ cells expressed GABA in the hippocampus and surrounding structures (n = 4; Fig. 6A–I). Confocal microscopy images showed that BrdU+ cells co-localized with GABA+ cells (Fig. 6J), and hNP+ cells dual-labeled with GABA had small bipolar neuronal processes (Fig. 6K–N). Most grafted cells, however, appeared not to show the morphological features of mature interneurons resembling host inhibitiory hippocampal interneurons. A few BrdU+ cells (∼3%) were found to express CALB2, a GABAergic interneuron subtype (Fig. 6O–Q). In the pilocarpine model, grafted NSPCs also differentiated into TUJ1+ neurons, OLIG2+ oligodendrocyte progenitors, and GFAP+ astrocytes, although a large number of cells expressed an undifferentiated cell marker, nestin (n = 4). Moreover, ∼21% of donor-derived cells expressed GABA and few of them expressed GDNF (Fig. S4). Most grafted cells also appeared not to show the morphological features of mature interneurons. These findings suggest that huNSPCs can give rise to neurons including GABAeric neurons albeit still immature, oligodendrocytes, and astrocytes in the epileptic hippocampus and adjacent structures in TLE models.


Human fetal brain-derived neural stem/progenitor cells grafted into the adult epileptic brain restrain seizures in rat models of temporal lobe epilepsy.

Lee H, Yun S, Kim IS, Lee IS, Shin JE, Park SC, Kim WJ, Park KI - PLoS ONE (2014)

Differentiation of human NSPCs following transplantation into the hippocampus of kindled rats.(A–F) BrdU+ grafted cells (green) were co-stained with TUJ1 (red, arrowheads in B, C, E, F) in the CA3 region of the hippocampus (A–C) and fimbria (D–F) in rats. (G–J) hNP+ grafted cells (red, arrowheads in G, I, J) were co-localized with TUJ1 (green, arrowheads in H–J) in the hilus of the hippocampus. (J) Orthogonal view from confocal z-series showed that hNP (red) in nuclei and TUJ1 (green) in cytoplasm were expressed in the same cell. (K, M) Under the dual-filter microscope, BrdU+ grafted cells co-expressed APC-CC1 in the fimbria (arrows in K), and hNuc+ grafted cells co-expressed GFAP in the CA3 region (an arrow in M). Scale bar; 200 µm (A), 20 µm (D), 10 µm (G, J), 20 µm (K).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4126719&req=5

pone-0104092-g005: Differentiation of human NSPCs following transplantation into the hippocampus of kindled rats.(A–F) BrdU+ grafted cells (green) were co-stained with TUJ1 (red, arrowheads in B, C, E, F) in the CA3 region of the hippocampus (A–C) and fimbria (D–F) in rats. (G–J) hNP+ grafted cells (red, arrowheads in G, I, J) were co-localized with TUJ1 (green, arrowheads in H–J) in the hilus of the hippocampus. (J) Orthogonal view from confocal z-series showed that hNP (red) in nuclei and TUJ1 (green) in cytoplasm were expressed in the same cell. (K, M) Under the dual-filter microscope, BrdU+ grafted cells co-expressed APC-CC1 in the fimbria (arrows in K), and hNuc+ grafted cells co-expressed GFAP in the CA3 region (an arrow in M). Scale bar; 200 µm (A), 20 µm (D), 10 µm (G, J), 20 µm (K).
Mentions: To study differentiation patterns of donor-derived cells, the whole hippocampal area of kindled rats was analyzed at 8 weeks following transplantation. BrdU+ cells were co-labeled with the early neuronal marker TUJ1 in the CA3 region of the hippocampus (Fig. 5A–C) and fimbria (Fig. 5D–F). Confocal microscopic images of hNP+-grafted cells demonstrated dual labeling with the TUJ1 antibody (Fig. 5G–J). Quantification revealed that donor-derived cells differentiated into TUJ1+ neurons (33.8±4.0%, n = 4), APC-CC1-expressing oligodendrocytes (27.7±6.2%, n = 4), and GFAP-expressing astrocytes (8.2±1.8%, n = 4) in the hippocampus (Fig. 5K, M). About 40% of GFAP-expressing astrocytes expressed GDNF (Fig. S3). Additionally, ∼24% of BrdU+ cells expressed GABA in the hippocampus and surrounding structures (n = 4; Fig. 6A–I). Confocal microscopy images showed that BrdU+ cells co-localized with GABA+ cells (Fig. 6J), and hNP+ cells dual-labeled with GABA had small bipolar neuronal processes (Fig. 6K–N). Most grafted cells, however, appeared not to show the morphological features of mature interneurons resembling host inhibitiory hippocampal interneurons. A few BrdU+ cells (∼3%) were found to express CALB2, a GABAergic interneuron subtype (Fig. 6O–Q). In the pilocarpine model, grafted NSPCs also differentiated into TUJ1+ neurons, OLIG2+ oligodendrocyte progenitors, and GFAP+ astrocytes, although a large number of cells expressed an undifferentiated cell marker, nestin (n = 4). Moreover, ∼21% of donor-derived cells expressed GABA and few of them expressed GDNF (Fig. S4). Most grafted cells also appeared not to show the morphological features of mature interneurons. These findings suggest that huNSPCs can give rise to neurons including GABAeric neurons albeit still immature, oligodendrocytes, and astrocytes in the epileptic hippocampus and adjacent structures in TLE models.

Bottom Line: However, NSPC grafting neither improved spatial learning or memory function in pilocarpine-treated animals.Grafted cells restored the expression of GDNF in host astrocytes but did not reverse the mossy fiber sprouting, eliminating the latter as potential mechanism.These results suggest that human fetal brain-derived NSPCs possess some therapeutic effect for TLE treatments although further studies to both increase the yield of NSPC grafts-derived functionally integrated GABAergic neurons and improve cognitive deficits are still needed.

View Article: PubMed Central - PubMed

Affiliation: Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT
Cell transplantation has been suggested as an alternative therapy for temporal lobe epilepsy (TLE) because this can suppress spontaneous recurrent seizures in animal models. To evaluate the therapeutic potential of human neural stem/progenitor cells (huNSPCs) for treating TLE, we transplanted huNSPCs, derived from an aborted fetal telencephalon at 13 weeks of gestation and expanded in culture as neurospheres over a long time period, into the epileptic hippocampus of fully kindled and pilocarpine-treated adult rats exhibiting TLE. In vitro, huNSPCs not only produced all three central nervous system neural cell types, but also differentiated into ganglionic eminences-derived γ-aminobutyric acid (GABA)-ergic interneurons and released GABA in response to the depolarization induced by a high K+ medium. NSPC grafting reduced behavioral seizure duration, afterdischarge duration on electroencephalograms, and seizure stage in the kindling model, as well as the frequency and the duration of spontaneous recurrent motor seizures in pilocarpine-induced animals. However, NSPC grafting neither improved spatial learning or memory function in pilocarpine-treated animals. Following transplantation, grafted cells showed extensive migration around the injection site, robust engraftment, and long-term survival, along with differentiation into β-tubulin III+ neurons (∼34%), APC-CC1+ oligodendrocytes (∼28%), and GFAP+ astrocytes (∼8%). Furthermore, among donor-derived cells, ∼24% produced GABA. Additionally, to explain the effect of seizure suppression after NSPC grafting, we examined the anticonvulsant glial cell-derived neurotrophic factor (GDNF) levels in host hippocampal astrocytes and mossy fiber sprouting into the supragranular layer of the dentate gyrus in the epileptic brain. Grafted cells restored the expression of GDNF in host astrocytes but did not reverse the mossy fiber sprouting, eliminating the latter as potential mechanism. These results suggest that human fetal brain-derived NSPCs possess some therapeutic effect for TLE treatments although further studies to both increase the yield of NSPC grafts-derived functionally integrated GABAergic neurons and improve cognitive deficits are still needed.

Show MeSH
Related in: MedlinePlus