Zebrafish (Danio rerio) is an emerged model used to study human disorders and cancer as it shares around 70 % of homologous genes with humans (Howe K, et al. 2013). Zebrafish can produce more than 100 embryos per clutch, for which the majority of organs are developed at 24 hours post fertilization. They remain transparent at early stage of their life cycle, enabling non- invasive imaging. Immaturity of their immune system, until day 7, allows or enables engraftment of human cell lines or patient-derived samples (Deveau et al, 2017)
Our previous objectives (described in the deliverable 2.1) were to achieve the standardization and the automation of the xenotransplantation process, the certification of our laboratories, and the dissemination activity and materials to relevant stakeholders. The purpose of the deliverable 2.2 is to report the work implemented for automation of the process using the cancer cell lines.
This automation is suitable for transparency of the procedure, identification of the troubleshooting points, minimization of costs and calculation of the business process (processing capability).
The following steps had been computerized:
– Fish breeding and egg collection: optimization of fish breeding
– In vitro:
- cancer cell culture
- determination of cell sensitivity to FDA-approved drugs • cell fluorescent staining
– In vivo:
- FDA-approved drug toxicity on zebrafish embryos
- xenotransplantation of zebrafish embryos
– Cancer development monitorization: placement of the xenograft embryos in an appropriated support for imaging by the Cytation5TM, thus providing high quality information about localization of cancerous cells inside the embryo.
– Data analysis: image extraction and analysis of cancer cells progression before and after specific drug administration.