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Teratoma Assay for Testing Pluripotency and Malignancy of Stem Cells: Insufficient Reporting and Uptake of Animal-Free Methods-A Systematic Review

Pluripotency describes the ability of stem cells to differentiate into derivatives of the three germ layers. In reporting new human pluripotent stem cell lines, their clonal derivatives or the safety of differentiated derivatives for transplantation, assessment of pluripotency is essential. Historic...

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Published in:International journal of molecular sciences 2023-02, Vol.24 (4), p.3879
Main Authors: Montilla-Rojo, Joaquin, Bialecka, Monika, Wever, Kimberley E, Mummery, Christine L, Looijenga, Leendert H J, Roelen, Bernard A J, Salvatori, Daniela C F
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description Pluripotency describes the ability of stem cells to differentiate into derivatives of the three germ layers. In reporting new human pluripotent stem cell lines, their clonal derivatives or the safety of differentiated derivatives for transplantation, assessment of pluripotency is essential. Historically, the ability to form teratomas in vivo containing different somatic cell types following injection into immunodeficient mice has been regarded as functional evidence of pluripotency. In addition, the teratomas formed can be analyzed for the presence of malignant cells. However, use of this assay has been subject to scrutiny for ethical reasons on animal use and due to the lack of standardization in how it is used, therefore questioning its accuracy. In vitro alternatives for assessing pluripotency have been developed such as ScoreCard and PluriTest. However, it is unknown whether this has resulted in reduced use of the teratoma assay. Here, we systematically reviewed how the teratoma assay was reported in publications between 1998 (when the first human embryonic stem cell line was described) and 2021. Our analysis of >400 publications showed that in contrast to expectations, reporting of the teratoma assay has not improved: methods are not yet standardized, and malignancy was examined in only a relatively small percentage of assays. In addition, its use has not decreased since the implementation of the ARRIVE guidelines on reduction of animal use (2010) or the introduction of ScoreCard (2015) and PluriTest (2011). The teratoma assay is still the preferred method to assess the presence of undifferentiated cells in a differentiated cell product for transplantation since the in vitro assays alone are not generally accepted by the regulatory authorities for safety assessment. This highlights the remaining need for an in vitro assay to test malignancy of stem cells.
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subjects Analysis
Animals
Assaying
Cancer
Cell Differentiation
Cell Line
Cell lines
Embryo cells
Embryonic stem cells
Embryonic Stem Cells - metabolism
Embryos
Ethical standards
hPSCs
Humans
Immunodeficiency
In vitro methods and tests
In vivo methods and tests
Injections
Laboratory animals
Malignancy
Mice
Pluripotency
Pluripotent Stem Cells - metabolism
Regulatory agencies
Review
Standardization
Stem cells
Systematic review
Teratoma
Teratoma - pathology
teratoma assay
Transplantation
Tumors
title Teratoma Assay for Testing Pluripotency and Malignancy of Stem Cells: Insufficient Reporting and Uptake of Animal-Free Methods-A Systematic Review
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