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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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Transcript expression profiling of human NSPCs.(A–F) Expression patterns of genes associated with development and function of GABAergic neurons were analyzed in NSPCs under proliferation (Prol) and differentiation conditions (Diff) using quantitative RT-PCR. The expression levels of each mRNA expression were normalized to levels of GAPDH. Panels show: telencephalic (A), ventral telencephalic GABAergic neuronal lineage (B), medial ganglionic eminence (MGE) (C), caudal ganglionic eminence (CGE) (D), GABAergic neuron (E), and interneuron (IN) subtype markers (F). The expression levels of makers were significantly elevated under Diff condition compared to under Prol condition except OLIG2. Data represented as mean ± SEM (n = 3).
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pone-0104092-g002: Transcript expression profiling of human NSPCs.(A–F) Expression patterns of genes associated with development and function of GABAergic neurons were analyzed in NSPCs under proliferation (Prol) and differentiation conditions (Diff) using quantitative RT-PCR. The expression levels of each mRNA expression were normalized to levels of GAPDH. Panels show: telencephalic (A), ventral telencephalic GABAergic neuronal lineage (B), medial ganglionic eminence (MGE) (C), caudal ganglionic eminence (CGE) (D), GABAergic neuron (E), and interneuron (IN) subtype markers (F). The expression levels of makers were significantly elevated under Diff condition compared to under Prol condition except OLIG2. Data represented as mean ± SEM (n = 3).

Mentions: We analyzed transcript expression for region-specific and GABAergic interneuron lineage markers in huNSPCs using qRT-PCR (Fig. 2). Telencephalic marker FOXG1 and ventral telencephalic markers (OLIG2, ASCL1, and DLX2) were expressed in cells under both Prol and Diff conditions [26], [41], and the expression levels of makers were significantly elevated under Diff condition compared to under Prol condition except OLIG2 (Fig. 2A, B). The MGE, marked by NKX2.1 and LHX6 expression, is known the primary source of cortical interneurons both in rodents and in humans, and the caudal ganglionic eminence (CGE), marked by NR2F2 (COUP-TFII) expression, give rise to a greater proportion of cortical interneurons in humans than in rodents [26], [42]-[45]. We found that NSPCs not only expressed MGE markers, but also abundantly the CGE marker (Fig. 2C, D). In addition, GABAergic markers (GAD1, SLC32A1, and SLC6A1) and interneuron subtype markers (CALB2, SST, and NPY) were robustly expressed under differentiation conditions (Fig. 2E, F). These data demonstrate that huNSPCs not only give rise to all three CNS neural cell types, but also that many of them differentiate into the MGE and CGE-derived GABAergic interneurons.


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)

Transcript expression profiling of human NSPCs.(A–F) Expression patterns of genes associated with development and function of GABAergic neurons were analyzed in NSPCs under proliferation (Prol) and differentiation conditions (Diff) using quantitative RT-PCR. The expression levels of each mRNA expression were normalized to levels of GAPDH. Panels show: telencephalic (A), ventral telencephalic GABAergic neuronal lineage (B), medial ganglionic eminence (MGE) (C), caudal ganglionic eminence (CGE) (D), GABAergic neuron (E), and interneuron (IN) subtype markers (F). The expression levels of makers were significantly elevated under Diff condition compared to under Prol condition except OLIG2. Data represented as mean ± SEM (n = 3).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126719&req=5

pone-0104092-g002: Transcript expression profiling of human NSPCs.(A–F) Expression patterns of genes associated with development and function of GABAergic neurons were analyzed in NSPCs under proliferation (Prol) and differentiation conditions (Diff) using quantitative RT-PCR. The expression levels of each mRNA expression were normalized to levels of GAPDH. Panels show: telencephalic (A), ventral telencephalic GABAergic neuronal lineage (B), medial ganglionic eminence (MGE) (C), caudal ganglionic eminence (CGE) (D), GABAergic neuron (E), and interneuron (IN) subtype markers (F). The expression levels of makers were significantly elevated under Diff condition compared to under Prol condition except OLIG2. Data represented as mean ± SEM (n = 3).
Mentions: We analyzed transcript expression for region-specific and GABAergic interneuron lineage markers in huNSPCs using qRT-PCR (Fig. 2). Telencephalic marker FOXG1 and ventral telencephalic markers (OLIG2, ASCL1, and DLX2) were expressed in cells under both Prol and Diff conditions [26], [41], and the expression levels of makers were significantly elevated under Diff condition compared to under Prol condition except OLIG2 (Fig. 2A, B). The MGE, marked by NKX2.1 and LHX6 expression, is known the primary source of cortical interneurons both in rodents and in humans, and the caudal ganglionic eminence (CGE), marked by NR2F2 (COUP-TFII) expression, give rise to a greater proportion of cortical interneurons in humans than in rodents [26], [42]-[45]. We found that NSPCs not only expressed MGE markers, but also abundantly the CGE marker (Fig. 2C, D). In addition, GABAergic markers (GAD1, SLC32A1, and SLC6A1) and interneuron subtype markers (CALB2, SST, and NPY) were robustly expressed under differentiation conditions (Fig. 2E, F). These data demonstrate that huNSPCs not only give rise to all three CNS neural cell types, but also that many of them differentiate into the MGE and CGE-derived GABAergic interneurons.

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