RESUMO
The hanging drop cell culture technique allows to study three-dimensional growth and differentiation of cell aggregates, that is, embryonic stem cells. Compared to standard two-dimensional monolayer cell cultivation, hanging drops allow for a better visualization and understanding of the developmental processes in vitro. Hanging drop cultivation can also be used to study biology of cancer cells three-dimensionally in vitro. This method can serve as an intermediate between the two-dimensional monolayer cell culture and in vivo models, which can be simply established in laboratories exhibiting minimum requirements of cell culture equipment. In this chapter, we describe the three-dimensional cultivation of germ cell cancer cell lines in hanging drops.
Assuntos
Técnicas de Cultura de Células , Neoplasias Embrionárias de Células Germinativas/patologia , Esferoides Celulares , Técnicas de Cultura de Células/métodos , Linhagem Celular Tumoral , HumanosRESUMO
Cisplatin resistance still remains a major obstacle in the standard chemotherapeutic approach in late-stage and metastatic testicular germ cell cancer (GCC) patients. This multifactorial and complex phenomenon arises (concomitantly) on several levels due to impaired transport, decreased adduct formation, increased DNA-repair, decreased apoptosis, or compensating pathways. Evaluation of novel therapeutic approaches and pharmacological inhibitors still remains necessary to treat cisplatin-resistant GCCs. In this chapter, we present in vitro techniques to measure cytotoxic impacts of chemotherapeutic drugs on GCC cell lines. Specifically, we will discuss the measurement of relative cell viability by XTT assay, as well as cell cycle distribution and apoptosis assay by Nicoletti- and Annexin V/PI apoptosis assay with subsequent flow cytometry, respectively, to evaluate the effects of cytotoxic treatment in GCC cell lines.