Selective Removal of Undifferentiated Embryonic Stem Cells from Differentiation Cultures Through HSV1 Thymidine Kinase and Ganciclovir Treatment

Naujok, Ortwin; Kaldrack, Joanna; Terbish, Taivankhuu; Jörns, Anne; Lenzen, Sigurd

doi:10.1007/s12015-010-9148-z

Cited by 34 publications

(25 citation statements)

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“…To achieve this several options have been considered: implantation of mature cells without contaminating residual undifferentiated cells, sorting techniques, positive selection utilizing resistance genes 16,17,102,103 and selective ablation of undifferentiated cells. [104][105][106] Thus, effective techniques to remove potentially teratogenic cells from mixed populations have been reported, but they have not yet been combined with protocols for differentiation of pluripotent cells into insulin-producing β-cells. This prevents pluripotent cells from clinical use at present.…”

Section: Patient-specific Stem Cellsmentioning

confidence: 99%

Insulin-producing Surrogate β-cells From Embryonic Stem Cells: Are We There Yet?

et al. 2011

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Embryonic stem cells (ESCs) harbor the potential to generate every cell type of the body by differentiation. The use of hESCs holds great promise for potential cell replacement therapies for degenerative diseases including diabetes mellitus. The recently discovered induced pluripotent stem cells (iPSCs) exhibit immense potential for regenerative medicine as they allow the generation of autologous cells tailored to the patients' immune system. Research for insulin-producing surrogate cells from ESCs has yielded highly controversial results, because many steps and factors in the differentiation process are currently still unknown. Thus, there is no consensus on common standard protocols. The protocols presently used established the differentiation from pluripotent cells toward pancreatic progenitor cells. However, none of the differentiation protocols reported to date have generated by exclusive in vitro differentiation sufficient numbers of insulin-producing cells meeting all essential criteria of a β-cell. The cells often lack the crucial function of regulated insulin secretion upon glucose stimulation. This review focuses on past and current approaches to the generation of insulin-producing cells from pluripotent sources, such as ESCs and iPSCs, and critically discusses the hurdles to be taken before insulin-secreting surrogate cells derived from these stem cells will be of clinical use in humans.

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Section: Patient-specific Stem Cellsmentioning

confidence: 99%

Insulin-producing Surrogate β-cells From Embryonic Stem Cells: Are We There Yet?

et al. 2011

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“…Therefore, eliminating the residual USCs is an indispensable step for safer cell therapy and efficient hepatotoxicity screening of drug candidates. Recently, selective removal of USCs has been tried to prevent tumorigenic potential and teratoma formation using antibody-based strategy in clinical fields [14–16]. In addition, small molecules have been suggested to effectively eliminate the risk of teratomas [17, 18].…”

Section: Introductionmentioning

confidence: 99%

Hepatic population derived from human pluripotent stem cells is effectively increased by selective removal of undifferentiated stem cells using YM155

et al. 2017

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BackgroundPluripotent stem cells (PSCs) such as embryonic stem cells and induced pluripotent stem cells are promising target cells for cell regenerative medicine together with recently advanced technology of in-vitro differentiation. However, residual undifferentiated stem cells (USCs) during in-vitro differentiation are considered a potential risk for development of cancer cells and nonspecific lineage cell types. In this study we observed that USCs still exist during hepatic differentiation, consequently resulting in poor quality of the hepatic population and forming teratoma in vivo. Therefore, we hypothesized that effectively removing USCs from in-vitro differentiation could improve the quality of the hepatic population and guarantee safety from risk of teratoma formation.MethodsHuman PSCs were differentiated to hepatocytes via four steps. YM155, a known BIRC5 inhibitor, was applied for removing the residual USCs on the hepatic differentiation. After YM155 treatment, hepatocyte development was evaluated by measuring gene expression, immunostaining and hepatic functions at each stage of differentiation, and forming teratomas were confirmed by cell transplantation with or without YM155.ResultsThe selected concentrations of YM155 removed USCs (NANOG+ and OCT4+) in a dose-dependent manner. As a result, expression of endodermal markers (SOX17, FOXA2 and CXCR4) at stage II of differentiation and hepatic markers (ALB, AFP and HNF4A) at stage III was up-regulated by YM155 treatment as well as the hepatic population (ALB+), and functions (ALB/urea secretion and CYP450 enzyme activity) were enhanced at the final stage of differentiation (stage IV). Furthermore, we demonstrated that NANOG and OCT4 expression remaining until stage III (day 15 of differentiation) completely disappeared when treated with YM155 and teratoma formation was effectively prevented by YM155 pretreatment in the in-vitro study.ConclusionsWe suggest that the removal of USCs using YM155 could improve the quantity and quality of induced hepatocytes and eliminate the potential risk of teratoma formation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0517-2) contains supplementary material, which is available to authorized users.

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“…This approach has been applied clinically and demonstrated efficacy in abrogating graft-versus-host disease in patients undergoing transplantation of donor T-cells modified with an inducible Caspase-9 gene [2]. Naujok et al have adapted this approach to PSCs through transduction of a construct where an OCT4 promoter controls HSV1 thymidine kinase expression, allowing for selective ablation of undifferentiated PSCs upon treatment with of Ganciclovir [3]. Unfortunately, these types of removal will eliminate the desired grafted cells along with teratoma-forming cells.…”

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confidence: 99%

The Safety of Embryonic Stem Cell Therapy Relies on Teratoma Removal

Tang

Drukker²

2012

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Selective Removal of Undifferentiated Embryonic Stem Cells from Differentiation Cultures Through HSV1 Thymidine Kinase and Ganciclovir Treatment

Cited by 34 publications

References 31 publications

Insulin-producing Surrogate β-cells From Embryonic Stem Cells: Are We There Yet?

Insulin-producing Surrogate β-cells From Embryonic Stem Cells: Are We There Yet?

Hepatic population derived from human pluripotent stem cells is effectively increased by selective removal of undifferentiated stem cells using YM155

The Safety of Embryonic Stem Cell Therapy Relies on Teratoma Removal

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