Disodium Cromolyn and Anti-podoplanin Antibodies Strongly Inhibit Growth of BHK 21/C13-derived Fibrosarcoma in a Chick Embryo Chorioallantoic Membrane Model

Abstract

Abstract. Aim: To characterize baby hamster kidney fibroblast (BHK 21/C13) cells and test the effects of antibodies against podoplanin and disodium cromolyn on BHK 21/C13 cell line-derived tumors grown on chick embryo chorioallantoic membrane (CAM). Material and MethodsExperimental models are still powerful tools for medical research. In vitro and in vivo models are designed to help researchers in their work for understanding disease mechanisms and for the discovery and testing of new therapeutics which by their future clinical application may improve the prognosis and survival of patients with various diseases (1-3). Despite researchers' efforts to create a perfect experimental model for each disease, several issues remained unresolved (4). Ethics rules become more and more strict regarding the use of animals in experimental models (5, 6) and thus the development of alternative experimental models which lack the use of animals is a real challenge for the scientific world. Even though in the future microfluidic systems such as tumor/organ on a chip model (7, 8) or 3D-printed organs may be used to develop new experimental models (9, 10), cell lines will remain the main component of any experimental model. Countless options for the use of cell lines are available today. There are normal and tumor cell lines used for various purposes from vaccine production (11) to testing cell toxicity of different agents (12, 13).Baby hamster kidney fibroblasts (BHK-21/C13) is a wellknown cell line sensitive to various viruses such as herpesvirus (14), hepatic (15) or rabies virus (16), but not polyoma virus. These cells are able to undergo malignant transformation, with fast growing behavior due to rapid proliferation and invasion of adjacent tissues when they are subcutaneously injected into hamsters. A giant fibrosarcomalike tumor without evidence of lymphatic or distant metastasis may develop at the site of inoculation. Few studies in the field of tumor experimental models used BHK-21/C13 cell line to obtain fibrosarcomas and even fewer to test the effects of different therapeutic agents.The BHK-21/C13 cell line has not been characterized regarding their phenotype except for some old articles which reported the expression of fibroblast growth factor by BHK-21/C13 cells (17) and the effects of vascular permeability factor on their proliferation and migration (18,19). Regarding BHK-21/C13 cell response to different therapeutic 791

Sarcoma treatment in the era of molecular medicine

“…In vivo , the chick chorioallantoic membrane (CAM) assay is a valuable option due to its low costs and relatively easy implementation. CAM assays have been employed to study sarcoma angiogenesis, fibroblast infiltration, tumorigenesis, tumor invasion, and metastasis in CHS, EwS, fibrosarcoma, LPS, and OS (Sys et al , 2013; Patil et al , 2014; Manjunathan & Ragunathan, 2015; Cimpean et al , 2018; Kunz et al , 2019; Perut et al , 2019; Steinestel et al , 2020). Numerous additional in vivo models of inducible or spontaneous sarcomas have been described in non‐mammalian vertebrates (e.g., zebrafish; Leacock et al , 2012; Mohseny et al , 2012; Brown et al , 2017b; Hayes & Langenau, 2017; Ignatius et al , 2018; Fleming et al , 2019) and in mammalians (e.g., mouse, rat, and dog; Cannon, 2015; Jacques et al , 2018; Castillo‐Tandazo et al , 2019; Pomella & Rota, 2020).…”

Hyaluronic Acid/Bone Substitute Complex Implanted on Chick Embryo Chorioallantoic Membrane Induces Osteoblastic Differentiation and Angiogenesis, but not Inflammation

“…Additionally, CAM is an embryonic tissue lacking an immune system and having mesenchymal cells with stem like potential able to differentiate depending on a specific microenvironment [15,16]. Based on the previously described features, CAM is a reliable in vivo model for testing the behavior of normal and pathologic tissues (as cultured cells or malignant tumors) [17,18], of different drugs and antibodies [19,20], or of a variety of biomaterials implanted on its surface [21,22]. There is minimal data available regarding bone implants on CAM [23,24].…”

Diagnostic Algorithm in Hirschsprung's Disease: Focus on Immunohistochemistry Markers

“…Staining for S-100 coupled with calretinin and PGP9.5 immunostaining performed on suction rectal biopsies was shown to be sensitive and specific for diagnosing HD (12). However, S-100 was found to be positive in ovarian cancer (13), metastases to the small intestine (14) and fibrosarcoma cells (15). The other marker, CD56, was used in evaluating a "transition zone" proximally to the aganglionic segment, with the use of serial colonic wall biopsies and obtained very good results (16).…”