{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T15:11:40Z","timestamp":1767625900053},"reference-count":32,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Neurosci"],"published-print":{"date-parts":[[2009,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>The human neural stem cell line CTX0E03 was developed for the cell based treatment of chronic stroke disability. Derived from fetal cortical brain tissue, CTX0E03 is a clonal cell line that contains a single copy of the c-mycER<jats:sup>TAM<\/jats:sup> transgene delivered by retroviral infection. Under the conditional regulation by 4-hydroxytamoxifen (4-OHT), c-mycER<jats:sup>TAM<\/jats:sup> enabled large-scale stable banking of the CTX0E03 cells. In this study, we investigated the fate of this transgene following growth arrest (EGF, bFGF and 4-OHT withdrawal) <jats:italic>in vitro<\/jats:italic> and following intracerebral implantation into a mid-cerebral artery occluded (MCAo) rat brain. <jats:italic>In vitro<\/jats:italic>, 4-weeks after removing growth factors and 4-OHT from the culture medium, c-mycER<jats:sup>TAM<\/jats:sup> transgene transcription is reduced by ~75%. Furthermore, immunocytochemistry and western blotting demonstrated a concurrent decrease in the c-MycER<jats:sup>TAM<\/jats:sup> protein. To examine the transcription of the transgene <jats:italic>in vivo<\/jats:italic>, CTX0E03 cells (450,000) were implanted 4-weeks post MCAo lesion and analysed for human cell survival and c-mycER<jats:sup>TAM<\/jats:sup> transcription by qPCR and qRT-PCR, respectively.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>The results show that CTX0E03 cells were present in all grafted animal brains ranging from 6.3% to 39.8% of the total cells injected. Prior to implantation, the CTX0E03 cell suspension contained 215.7 (SEM = 13.2) copies of the c-mycER<jats:sup>TAM<\/jats:sup> transcript per cell. After implantation the c-mycER<jats:sup>TAM<\/jats:sup> transcript copy number per CTX0E03 cell had reduced to 6.9 (SEM = 3.4) at 1-week and 7.7 (SEM = 2.5) at 4-weeks. Bisulfite genomic DNA sequencing of the <jats:italic>in vivo<\/jats:italic> samples confirmed c-mycER<jats:sup>TAM<\/jats:sup> silencing occurred through methylation of the transgene promoter sequence.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>In conclusion the results confirm that CTX0E03 cells downregulated c-mycER<jats:sup>TAM<\/jats:sup> transgene expression both <jats:italic>in vitro<\/jats:italic> following EGF, bFGF and 4-OHT withdrawal and <jats:italic>in vivo<\/jats:italic> following implantation in MCAo rat brain. The silencing of the c-mycER<jats:sup>TAM<\/jats:sup> transgene <jats:italic>in vivo<\/jats:italic> provides an additional safety feature of CTX0E03 cells for potential clinical application.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1471-2202-10-86","type":"journal-article","created":{"date-parts":[[2009,7,21]],"date-time":"2009-07-21T18:29:25Z","timestamp":1248200965000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["c-MycERTAM transgene silencing in a genetically modified human neural stem cell line implanted into MCAo rodent brain"],"prefix":"10.1186","volume":"10","author":[{"given":"Lara","family":"Stevanato","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Randolph L","family":"Corteling","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul","family":"Stroemer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew","family":"Hope","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julie","family":"Heward","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erik A","family":"Miljan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John D","family":"Sinden","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2009,7,21]]},"reference":[{"issue":"2","key":"1090_CR1","doi-asserted-by":"publisher","first-page":"140","DOI":"10.1016\/j.brainres.2005.09.049","volume":"1063","author":"AM Wong","year":"2005","unstructured":"Wong AM, Hodges H, Horsburgh K: Neural stem cell grafts reduce the extent of neuronal damage in a mouse model of global ischaemia. 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