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. 2022 Aug 1;7:38. doi: 10.1038/s41536-022-00234-7

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Status epilepticus (SE) was induced in two-month-old male F344 rats through graded kainic acid injections. Two months later, animals exhibiting chronic temporal lobe epilepsy (TLE) typified by spontaneous recurrent seizures (SRS) were chosen for bilateral grafting of human medial ganglionic eminence (hMGE)-like cells. The hMGE cells were generated from hM4Di-receptor (a designer receptor activated only by designer drugs, DREADDs) expressing human embryonic stem cell (hESC)-line through a directed differentiation protocol. In the fourth month after grafting, rats were implanted with electrodes, and 2 weeks later, continuous video-encephalographic (video-EEG) recordings were taken before (days 1–5), during (days 6–8), and 2 days after (days 11–14) clozapine-N-oxide (CNO) treatment. Seven days after completing video-EEG recordings, the animals were probed with the object location tests (OLTs), with and without CNO administration, to assess hippocampus-dependent cognitive function. Seven days after completing the second OLT, the animals were investigated with the pattern separation tests (PSTs), with and without CNO administration, to examine their pattern separation ability. Following behavioral tests, the animals were perfused, and fixed brain tissues were processed to analyze the differentiation and integration of graft-derived cells through dual and triple immunofluorescence methods and confocal imaging.