Marine Collagen Substrates for 2D and 3D Ovarian Cancer Cell Systems

Abstract

A fundamental structural component of extracellular matrix in all connective and interstitial tissue, collagen is the most abundant protein in the human body. To date, mammalian collagens sources represent the golden standard for multiple biomedical applications, while marine-derived collagens have largely been used in industry (food, pharmaceutical, and cosmetic), with little use in research and clinical applications. Herein we demonstrate the effective use Rhizostoma pulmo jellyfish collagen, a source of biocompatible, sustainable collagen for 2D and 3D cell culture, addressing the global drive for technological developments that result in the replacement of animals and their derived products in research. Jellyfish collagen harbors similar structural features mammalian collagen type I, despite differing slightly in amino acid content. Jellyfish collagen supports ovarian cancer (OvCa) cell line proliferation, cellular morphology and expression of epithelial to mesenchymal transition markers, supporting the use of R. pulmo as a non-mammalian collagen cell culture substrate. Furthermore, R. pulmo collagen is effective in 3D device fabrication such as sponges where it mimics tissue architecture complexity. OvCa cells migrated and differentiated within the R. pulmo collagen 3D scaffolds confirming its suitability for advanced cell culturing applications, providing an excellent alternative to mammalian collagen sources for the culture of human cells.

Production of Injectable Marine Collagen-Based Hydrogel for the Maintenance of Differentiated Chondrocytes in Tissue Engineering Applications

“…The R. pulmo acid-soluble proteins were purified a Under reducing conditions, the acid-soluble proteins showed a pattern mainly consisting in α1 and α2 chains (approximately 180 kDa), and the β and γ chains located on a high molecular mass region (above 240 kDa), with a pattern likely resembling type I and type II collagen. This results in accordance with data on collagen isolated from several marine sources [36,38]. Additionally, the highly resolved protein pattern indicates that the used protocol allows the extraction of jellyfish Under reducing conditions, the acid-soluble proteins showed a pattern mainly consisting in α1 and α2 chains (approximately 180 kDa), and the β and γ chains located on a high molecular mass region (above 240 kDa), with a pattern likely resembling type I and type II collagen.…”

“…Additionally, the highly resolved protein pattern indicates that the used protocol allows the extraction of jellyfish collagen in native form. Although it is usually indicated a high correspondence between the collagen of mammalian and marine species [36], the apparent molecular weight of R. pulmo α and β chains was higher than the vertebrate counterparts. This could indicate variations in protein length and structural differences, probably due to their different amino acid composition.…”

“…This could indicate variations in protein length and structural differences, probably due to their different amino acid composition. In this line, it has been observed that R. pulmo collagen contains less hydroxyproline, proline, glycine and glutamic acid residues than rat and bovine type I collagen [36]. Whilst these amino acids are involved in the formation and stability of the quaternary structure (triple helix) and thermal stability of type I collagen, the reduced content of these amino acids did not appear to affect the structure of R. pulmo collagen [40].…”

The Biological Evaluation of Jellyfish Collagen as a New Research Tool for the Growth and Culture of iPSC Derived Microglia

“…Viability of cells was confirmed visually by capturing stained images under phase contrast (Figure 1), as well as by quantifying ramification of iMGL using the InCucyte S3 TM an indicator of robust microglial morphology (Figure 5). Solid evidence from diverse cell culture studies have shown the potential of jellyfish collagen to successfully culture a wide range of human cell types (Paradiso et al, 2019). In this study, we have successfully demonstrated that jellyfish collagen can also be used as a substrate for the culture of human iPSC-derived Microglia (iMGL) that when cultured, display the morphological, surface marker expression and functional characteristics expected from microglia.…”

“…This makes JFC an interesting molecule for cell culture study. Previous reports have demonstrated the potential of JFC for the culture of human cells in vitro (Song et al, 2006;Addad et al, 2011;Paradiso et al, 2019). Jellyfish collagen when compared against mammalian fibrillar collagen in cell cytotoxicity and cell adhesion assays, showed no statistical difference in cytotoxicity.…”