{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T11:51:20Z","timestamp":1788609080451,"version":"build-2803163510"},"reference-count":47,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2021,12,8]],"date-time":"2021-12-08T00:00:00Z","timestamp":1638921600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000780","name":"European Commission","doi-asserted-by":"publisher","award":["722779"],"award-info":[{"award-number":["722779"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004359","name":"Swedish Research Council","doi-asserted-by":"publisher","award":["2017-00921"],"award-info":[{"award-number":["2017-00921"]}],"id":[{"id":"10.13039\/501100004359","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministry of Health, Czech Republic","award":["21-08-00533"],"award-info":[{"award-number":["21-08-00533"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Epilepsy is a complex disorder affecting the central nervous system and is characterised by spontaneously recurring seizures (SRSs). Epileptic patients undergo symptomatic pharmacological treatments, however, in 30% of cases, they are ineffective, mostly in patients with temporal lobe epilepsy. Therefore, there is a need for developing novel treatment strategies. Transplantation of cells releasing \u03b3-aminobutyric acid (GABA) could be used to counteract the imbalance between excitation and inhibition within epileptic neuronal networks. We generated GABAergic interneuron precursors from human embryonic stem cells (hESCs) and grafted them in the hippocampi of rats developing chronic SRSs after kainic acid-induced status epilepticus. Using whole-cell patch-clamp recordings, we characterised the maturation of the grafted cells into functional GABAergic interneurons in the host brain, and we confirmed the presence of functional inhibitory synaptic connections from grafted cells onto the host neurons. Moreover, optogenetic stimulation of grafted hESC-derived interneurons reduced the rate of epileptiform discharges in vitro. We also observed decreased SRS frequency and total time spent in SRSs in these animals in vivo as compared to non-grafted controls. These data represent a proof-of-concept that hESC-derived GABAergic neurons can exert a therapeutic effect on epileptic animals presumably through establishing inhibitory synapses with host neurons.<\/jats:p>","DOI":"10.3390\/ijms222413243","type":"journal-article","created":{"date-parts":[[2021,12,8]],"date-time":"2021-12-08T23:30:00Z","timestamp":1639006200000},"page":"13243","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Human Stem Cell-Derived GABAergic Interneurons Establish Efferent Synapses onto Host Neurons in Rat Epileptic Hippocampus and Inhibit Spontaneous Recurrent Seizures"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5577-1206","authenticated-orcid":false,"given":"Eli\u0161ka","family":"Waloschkov\u00e1","sequence":"first","affiliation":[{"name":"Epilepsy Center, Department of Clinical Sciences, Lund University Hospital, 221 84 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8569-5566","authenticated-orcid":false,"given":"Ana","family":"Gonzalez-Ramos","sequence":"additional","affiliation":[{"name":"Epilepsy Center, Department of Clinical Sciences, Lund University Hospital, 221 84 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3530-8895","authenticated-orcid":false,"given":"Apostolos","family":"Mikroulis","sequence":"additional","affiliation":[{"name":"Epilepsy Center, Department of Clinical Sciences, Lund University Hospital, 221 84 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5597-8709","authenticated-orcid":false,"given":"Jan","family":"Kudl\u00e1\u010dek","sequence":"additional","affiliation":[{"name":"Epilepsy Center, Department of Clinical Sciences, Lund University Hospital, 221 84 Lund, Sweden"},{"name":"Department of Physiology, Second Faculty of Medicine, Charles University, 150 06 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3699-138X","authenticated-orcid":false,"given":"My","family":"Andersson","sequence":"additional","affiliation":[{"name":"Epilepsy Center, Department of Clinical Sciences, Lund University Hospital, 221 84 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8317-7976","authenticated-orcid":false,"given":"Marco","family":"Ledri","sequence":"additional","affiliation":[{"name":"Epilepsy Center, Department of Clinical Sciences, Lund University Hospital, 221 84 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Merab","family":"Kokaia","sequence":"additional","affiliation":[{"name":"Epilepsy Center, Department of Clinical Sciences, Lund University Hospital, 221 84 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/S1474-4422(18)30454-X","article-title":"Global, regional, and national burden of epilepsy, 1990\u20132016: A systematic analysis for the Global Burden of Disease Study 2016","volume":"18","author":"Beghi","year":"2019","journal-title":"Lancet Neurol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1016\/j.braindev.2012.09.002","article-title":"Effects of intravenous diazepam on high-frequency oscillations in EEGs with CSWS","volume":"35","author":"Toda","year":"2013","journal-title":"Brain Dev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.neulet.2011.02.045","article-title":"Gabapentin promotes inhibition by enhancing hyperpolarization-activated cation currents and spontaneous firing in hippocampal CA1 interneurons","volume":"494","author":"Peng","year":"2011","journal-title":"Neurosci. Lett."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1124\/jpet.109.153908","article-title":"Topiramate reduces excitability in the basolateral amygdala by selectively inhibiting GluK1 (GluR5) kainate receptors on interneurons and positively modulating GABAA receptors on principal neurons","volume":"330","author":"Braga","year":"2009","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/0143-4179(91)90120-8","article-title":"Vigabatrin pre-treatment prevents hilar somatostatin cell loss and the development of interictal spiking activity following sustained simulation of the perforant path","volume":"19","author":"Ylinen","year":"1991","journal-title":"Neuropeptides"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1111\/epi.12300","article-title":"Proposal for a \u201cphase II\u201d multicenter trial model for preclinical new antiepilepsy therapy development","volume":"54","author":"Bertram","year":"2013","journal-title":"Epilepsia"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.yebeh.2017.08.039","article-title":"Psychiatric and behavioral side effects of antiepileptic drugs in adults with epilepsy","volume":"76","author":"Chen","year":"2017","journal-title":"Epilepsy Behav."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1133","DOI":"10.1111\/j.1528-1167.2005.01605.x","article-title":"Psychological Distress, Comorbidities, and Health Behaviors among U.S. Adults with Seizures: Results from the 2002 National Health Interview Survey","volume":"46","author":"Strine","year":"2005","journal-title":"Epilepsia"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1111\/j.1600-0404.2010.01440.x","article-title":"Non-interventional surveillance study of adverse events in patients with epilepsy","volume":"124","author":"Cramer","year":"2011","journal-title":"Acta Neurol. Scand."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.1016\/S0140-6736(06)68477-8","article-title":"Adult epilepsy","volume":"367","author":"Duncan","year":"2006","journal-title":"Lancet"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1046\/j.1528-1157.43.s.6.6.x","article-title":"Epilepsy in the world today: The medical point of view","volume":"43","author":"Engel","year":"2002","journal-title":"Epilepsia"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"781","DOI":"10.1016\/j.psc.2007.07.005","article-title":"Psychiatric Aspects of Epilepsy","volume":"30","author":"Marcangelo","year":"2007","journal-title":"Psychiatr. Clin. N. Am."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"922","DOI":"10.1001\/jama.2012.220","article-title":"Early surgical therapy for drug-resistant temporal lobe epilepsy: A randomized trial","volume":"307","author":"Engel","year":"2012","journal-title":"JAMA J. Am. Med. Assoc."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1093\/jnen\/59.10.907","article-title":"Subregional Pathology of the Amygdala Complex and Entorhinal Region in Surgical Specimens from Patients with Pharmacoresistant Temporal Lobe Epilepsy","volume":"59","author":"Wolf","year":"2000","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3948","DOI":"10.1152\/jn.01378.2005","article-title":"Interneuron and pyramidal cell interplay during in vitro seizure-like events","volume":"95","author":"Ziburkus","year":"2006","journal-title":"J. Neurophysiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.yebeh.2005.07.019","article-title":"Axonal sprouting of GABAergic interneurons in temporal lobe epilepsy","volume":"7","author":"Bausch","year":"2005","journal-title":"Epilepsy Behav."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1002\/(SICI)1098-1063(1996)6:4<347::AID-HIPO1>3.0.CO;2-I","article-title":"Interneurons of the hippocampus","volume":"6","author":"Freund","year":"1998","journal-title":"Hippocampus"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1007\/s12264-014-1478-4","article-title":"Dysfunction of hippocampal interneurons in epilepsy","volume":"30","author":"Liu","year":"2014","journal-title":"Neurosci. Bull."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"14247","DOI":"10.1523\/JNEUROSCI.3842-09.2009","article-title":"Surviving hilar somatostatin interneurons enlarge, sprout axons, and form new synapses with granule cells in a mouse model of temporal lobe epilepsy","volume":"29","author":"Zhang","year":"2009","journal-title":"J. Neurosci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"876","DOI":"10.1002\/cne.21207","article-title":"Selective loss of dentate hilar interneurons contributes to reduced synaptic inhibition of granule cells in an electrical stimulation-based animal model of temporal lobe epilepsy","volume":"500","author":"Sun","year":"2007","journal-title":"J. Comp. Neurol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"692","DOI":"10.1038\/nn.3392","article-title":"GABA progenitors grafted into the adult epileptic brain control seizures and abnormal behavior","volume":"16","author":"Hunt","year":"2013","journal-title":"Nat. Neurosci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1002\/stem.446","article-title":"Medial ganglionic eminence-derived neural stem cell grafts ease spontaneous seizures and restore GDNF expression in a rat model of chronic temporal lobe epilepsy","volume":"28","author":"Waldau","year":"2010","journal-title":"Stem Cells"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1002\/ana.25021","article-title":"Persistent seizure control in epileptic mice transplanted with gamma-aminobutyric acid progenitors","volume":"82","author":"Casalia","year":"2017","journal-title":"Ann. Neurol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"13492","DOI":"10.1523\/JNEUROSCI.0005-14.2014","article-title":"Long-Term Seizure Suppression and Optogenetic Analyses of Synaptic Connectivity in Epileptic Mice with Hippocampal Grafts of GABAergic Interneurons","volume":"34","author":"Henderson","year":"2014","journal-title":"J. Neurosci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"15472","DOI":"10.1073\/pnas.0900141106","article-title":"Reduction of seizures by transplantation of cortical GABAergic interneuron precursors into Kv1.1 mutant mice","volume":"106","author":"Baraban","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1016\/j.expneurol.2008.04.040","article-title":"Grafting of striatal precursor cells into hippocampus shortly after status epilepticus restrains chronic temporal lobe epilepsy","volume":"212","author":"Hattiangady","year":"2008","journal-title":"Exp. Neurol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1016\/j.stem.2014.10.006","article-title":"hPSC-derived maturing GABAergic interneurons ameliorate seizures and abnormal behavior in epileptic mice","volume":"15","author":"Cunningham","year":"2014","journal-title":"Cell Stem Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1073\/pnas.1814185115","article-title":"Human induced pluripotent stem cell-derived MGE cell grafting after status epilepticus attenuates chronic epilepsy and comorbidities via synaptic integration","volume":"116","author":"Upadhya","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","first-page":"1","article-title":"Human stem cell-derived GABAergic neurons functionally integrate into human neuronal networks","volume":"11","author":"Mikroulis","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1038\/nmeth.4291","article-title":"Generation of pure GABAergic neurons by transcription factor programming","volume":"14","author":"Yang","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2548","DOI":"10.1016\/j.stemcr.2021.08.006","article-title":"Defining the nature of human pluripotent stem cell-derived interneurons via single-cell analysis","volume":"16","author":"Allison","year":"2021","journal-title":"Stem Cell Rep."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Shrestha, S., Anderson, N.C., Grabel, L.B., Naegele, J.R., and Aaron, G.B. (2020). Development of electrophysiological and morphological properties of human embryonic stem cell-derived GABAergic interneurons at different times after transplantation into the mouse hippocampus. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0237426"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.scr.2018.10.007","article-title":"Pluripotent stem cell-derived interneuron progenitors mature and restore memory deficits but do not suppress seizures in the epileptic mouse brain","volume":"33","author":"Anderson","year":"2018","journal-title":"Stem Cell Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"8465","DOI":"10.1073\/pnas.0602979103","article-title":"Anomalous levels of Cl- transporters in the hippocampal subiculum from temporal lobe epilepsy patients make GABA excitatory","volume":"103","author":"Palma","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"9866","DOI":"10.1523\/JNEUROSCI.2761-07.2007","article-title":"Perturbed chloride homeostasis and GABAergic signaling in human temporal lobe epilepsy","volume":"27","author":"Huberfeld","year":"2007","journal-title":"J. Neurosci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"65","DOI":"10.3389\/fphar.2012.00065","article-title":"Excitatory GABA: How a correct observation may turn out to be an experimental artifact","volume":"3","author":"Bregestovski","year":"2012","journal-title":"Front. Pharmacol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/S0306-4522(99)00474-1","article-title":"Changes in the distribution and connectivity of interneurons in the epileptic human dentate gyrus","volume":"96","author":"Wittner","year":"2000","journal-title":"Neuroscience"},{"key":"ref_38","first-page":"100","article-title":"The vulnerability of calretinin-containing hippocampal interneurons to temporal lobe epilepsy","volume":"8","year":"2014","journal-title":"Front. Neuroanat."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2104","DOI":"10.1016\/j.stem.2021.09.002","article-title":"Reprogramming reactive glia into interneurons reduces chronic seizure activity in a mouse model of mesial temporal lobe epilepsy","volume":"28","author":"Lentini","year":"2021","journal-title":"Cell Stem Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1038\/nature05744","article-title":"Multimodal fast optical interrogation of neural circuitry","volume":"446","author":"Zhang","year":"2007","journal-title":"Nature"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"108542","DOI":"10.1016\/j.jneumeth.2019.108542","article-title":"Optimization of production and transgene expression of a retrogradely transported pseudotyped lentiviral vector","volume":"336","author":"Lockowandt","year":"2020","journal-title":"J. Neurosci. Methods"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/0013-4694(72)90177-0","article-title":"Modification of seizure activity by electrical stimulation: II. Motor seizure","volume":"32","author":"Racine","year":"1972","journal-title":"Electroencephalogr. Clin. Neurophysiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.nbd.2015.11.014","article-title":"Translational approach for gene therapy in epilepsy: Model system and unilateral overexpression of neuropeptide Y and Y2 receptors","volume":"86","author":"Melin","year":"2016","journal-title":"Neurobiol. Dis."},{"key":"ref_44","unstructured":"(2021, November 22). GitHub\u2014AMikroulis\/xPSC-Detection: Template Correlation-Based Detection of Postsynaptic Currents. Available online: https:\/\/github.com\/AMikroulis\/xPSC-detection."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1707","DOI":"10.1038\/sj.bjp.0701563","article-title":"The interaction of general anaesthetics with recombinant GABA(A) and glycine receptors expressed in Xenopus laevis oocytes: A comparative study","volume":"122","author":"Pistis","year":"1997","journal-title":"Br. J. Pharmacol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/S0028-3908(03)00032-7","article-title":"The GABAA receptor antagonist picrotoxin inhibits 5-hydroxytryptamine type 3A receptors","volume":"44","author":"Das","year":"2003","journal-title":"Neuropharmacology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1124\/jpet.104.080325","article-title":"Molecular determinants of picrotoxin inhibition of 5-hydroxytryptamine type 3 receptors","volume":"314","author":"Das","year":"2005","journal-title":"J. Pharmacol. Exp. Ther."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/24\/13243\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:43:45Z","timestamp":1760168625000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/24\/13243"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,8]]},"references-count":47,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["ijms222413243"],"URL":"https:\/\/doi.org\/10.3390\/ijms222413243","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,8]]}}}