AUTHOR=Merlos Rodrigo Miguel Angel , Casar Berta , Michalkova Hana , Jimenez Jimenez Ana Maria , Heger Zbynek , Adam Vojtech 

TITLE=Extending the Applicability of In Ovo and Ex Ovo Chicken Chorioallantoic Membrane Assays to Study Cytostatic Activity in Neuroblastoma Cells

JOURNAL=Frontiers in Oncology

VOLUME=11

YEAR=2021

URL=https://www.frontiersin.org/journals/oncology/articles/10.3389/fonc.2021.707366

DOI=10.3389/fonc.2021.707366

ISSN=2234-943X

ABSTRACT=<sec><title>Purpose</title><p>The chick chorioallantoic membrane (CAM) assay can provide an alternative versatile, cost-effective, and ethically less controversial <italic>in vivo</italic> model for reliable screening of drugs. In the presented work, we demonstrate that CAM assay (<italic>in ovo</italic> and <italic>ex ovo</italic>) can be simply employed to delineate the effects of cisplatin (CDDP) and ellipticine (Elli) on neuroblastoma (Nbl) cells in terms of their growth and metastatic potential.</p></sec><sec><title>Methods</title><p>The Nbl UKF-NB-4 cell line was established from recurrent bone marrow metastases of high-risk Nbl (stage IV, <italic>MYCN</italic> amplification, 7q21 gain). <italic>Ex ovo</italic> and <italic>in ovo</italic> CAM assays were optimized to evaluate the antimetastatic activity of CDDP and Elli. Immunohistochemistry, qRT-PCR, and DNA isolation were performed.</p></sec><sec><title>Results</title><p><italic>Ex ovo</italic> CAM assay was employed to study whether CDDP and Elli exhibit any inhibitory effects on growth of Nbl xenograft in <italic>ex ovo</italic> CAM assay. Under the optimal conditions, Elli and CDDP exhibited significant inhibition of the size of the primary tumor. To study the efficiency of CDDP and Elli to inhibit primary Nbl tumor growth, intravasation, and extravasation in the organs, we adapted the <italic>in ovo</italic> CAM assay protocol. In <italic>in ovo</italic> CAM assay, both studied compounds (CDDP and Elli) exhibited significant (<italic>p</italic> &lt; 0.001) inhibitory activity against extravasation to all investigated organs including distal CAM.</p></sec><sec><title>Conclusions</title><p>Taken together, CAM assay could be a helpful and highly efficient <italic>in vivo</italic> approach for high-throughput screening of libraries of compounds with expected anticancer activities.</p></sec>