Innovative organotypic in vitro models for safety assessment: aligning with regulatory requirements and understanding models of the heart, skin, and liver as paradigms

The development of improved, innovative models for the detection of toxicity of drugs, chemicals, or chemicals in cosmetics is crucial to efficiently bring new products safely to market in a cost-effective and timely manner. In addition, improvement in models to detect toxicity may reduce the incidence of unexpected post-marketing toxicity and reduce or eliminate the need for animal testing. The safety of novel products of the pharmaceutical, chemical, or cosmetics industry must be assured; therefore, toxicological properties need to be assessed. Accepted methods for gathering the information required by law for approval of substances are often animal methods. To reduce, refine, and replace animal testing, innovative organotypic in vitro models have emerged. Such models appear at different levels of complexity ranging from simpler, self-organized three-dimensional (3D) cell cultures up to more advanced scaffold-based co-cultures consisting of multiple cell types. This review provides an overview of recent developments in the field of toxicity testing with in vitro models for three major organ types: heart, skin, and liver. This review also examines regulatory aspects of such models in Europe and the UK, and summarizes best practices to facilitate the acceptance and appropriate use of advanced in vitro models

In vivo tumorigenicity of the 20q11.21 amplicon in an engraftment model of hPSCs and differentiated liver cells

Human pluripotent stem cells (hPSCs) are a promising source of somatic cells for clinical applications and disease modelling. However, during culture they accumulate genetic aberrations such as amplification of 20q11.21 which occurs in approximately 20% of extensively cultured hPSC lines and confers a BCL2L1-mediated survival advantage. During the production of the large number of cells required for transplantation and therapy these aberrations may become unavoidable which has important safety implications for therapies and may also impact upon disease modelling. Presently, these risks are poorly understood; whilst it is apparent that large-scale genetic aberrations can pose an oncogenic risk, the risks associated with smaller, more insidious changes have not been fully explored. In this report, the effects of engraftment of human embryonic stem cells (hESCs) and hESC-derived hepatocyte-like cells (HLCs) with and without amplification of the 20q11.21 minimal amplicon and isochromosome 20q (i20q) in SCID-beige mice are presented. The cells were tracked in vivo using a luminescent reporter over a period of approximately four months. Intrasplenic injection of hESCs showed greater engraftment potential and the formation of more severely disruptive lesions in the liver and spleen of animals injected with cells containing 20q11.21 compared with i20q and wild type. HLCs with 20q11.21 engrafted more successfully and formed more severely disruptive lesions than wild type cells or cells with i20q. These results reinforce the notion that karyotyping of therapeutic hPSC is required for transplant, and suggest that screening for known common aberrations is necessary. Further work to identify commonly arising genetic aberrations should be performed and routine screening for hPSCs intended for therapeutic use should be used

Longevidade de inflorescências de Epidendrum ibaguense tratadas com aminoetoxivinilglicina Extending vase life of cut Epidendrum ibaguense inflorescences with aminoethoxyvinylglycine

O objetivo deste trabalho foi avaliar a influência do inibidor da síntese de etileno aminoetoxivinilglicina (AVG) aplicado na forma de solução de condicionamento e pulverização sobre a abscisão e longevidade de inflorescências de Epidendrum ibaguense Kunth. As hastes foram colhidas e imediatamente condicionadas em solução com 0, 0,5, 1, 1,5 e 2 mM de AVG por 6, 12, 18 e 24 horas. Os mesmos tratamentos foram aplicados na forma de pulverização até o molhamento completo da inflorescência. O experimento foi conduzido em um esquema fatorial entre tempo de aplicação e doses de AVG, mais doses de AVG aplicadas em pulverização nas flores, e o delineamento foi em blocos ao acaso, com cinco repetições, com três hastes por unidade experimental. Independentemente do modo de aplicação do AVG, as concentrações utilizadas promoveram aumento da longevidade das flores em aproximadamente 70% em comparação ao controle, com resposta máxima nas concentrações de 1,5 e 2 mM. Apercentagem de abscisão de flores foi reduzida em todos os tratamentos com AVG, principalmente quando se utilizou pulverização, com decréscimo na abscisão acumulada superior a 80% nas concentrações entre 1 e 2 mM de AVG.A aplicação de AVG prolongaa longevidadee reduz a abscisãode flores de Epidendrum ibaguense.<br>This work evaluated the influence of the aminoethoxyvinylglycine (AVG), an inhibitor of ethylene synthesis, when applied in pulsing solution or sprayed, on the abscission and longevity of cut star orchid (Epidendrum ibaguense Kunth) inflorescences. The cut stems were placed in solutions with 0, 0.5, 1, 1.5 and 2 mM AVG concentrations immediately after the harvest for periods of 6, 12, 18 and 24 hours. The same treatment concentrations were sprayed on the inflorescences until runoff. The experiment had a randomized complete block design with factorial treatments of rate and duration plus rate of application to the flowers, with five replicates, with three stems per experimental unit. Regardless of the way the AVG was applied, the inhibitor extended the flowers'longevity in approximately 70% compared to the control treatment, reaching maximum effect at 1.5 and 2 mM AVG. The percentage of abscissed flowers diminished in all treatments containing AVG, with higher efficiency when sprayed over the inflorescence, decreasing the abscission by 80% at concentrations between 1 and 2 mM AVG