Antibiotics and antimycotics are currently indispensable for primary cell cultures of human origin, as microbial charge accumulated during biopsy collection would compromise cell survival. It is assumed that their presence in culture media does not significantly affect diagnostic tests or experimental protocols. However, antibiotics can activate intracellular pathways influencing cellular metabolism and signaling responses. Moreover, demanding cultures of gene-edited cells often suffer from poor viability rates, a limitation worsened by antibiotics. At the same time, the wide use of antibiotics and antimycotics contributes to pollutant accumulation.

In this context, the CNR-ISSMC team proposes a new antimicrobial material based on chitosan and copper ions (Cu²⁺), capable of minimizing the risk of bacterial or mycotic infections in cell culture plates, even in the absence of antibiotic solutions.

These customizable 2D and 3D porous matrices are:

• tailored for different cell cultures
• designed following a Safe and Sustainable-by-Design (SSbD) approach
• produced using raw materials derived from food chain waste (gelatin, chitosan)
• enriched with Cu²⁺-doped calcium phosphate (CaP) nanoparticles, synthesized through green manufacturing approaches

The material will be embedded in cell culture plates and slides and three highly qualified research institutes — CNR-IGM (expert in 2D primary human cell cultures), UNIMORE (expert in gene-editing and T cells manipulation), and CNR-IFT (expert in 3D cell cultures and organoids) — will assess the kit and standardize the conditions for antibiotic-free culturing.

Expected improvements include:

• higher quality of primary cell cultures such as pluripotent stem cells, bone-derived mesenchymal stem cells and osteoblasts, skin-derived tissue equivalents, adipose tissue stem cells, muscle-derived myoblasts and fibro-adipogenic precursors, lung-derived epithelial cells and organoids, and T cell cultures
• improved viability rate in cultures of gene-edited cells from patients with genetic diseases, supporting the development of advanced gene therapies (UNIMORE)
• reduction of pollution by limiting antibiotic use
• more efficient conditions for cell culturing
• increased interoperability among laboratories
• reduced cytotoxicity in advanced therapies such as those based on genome editing

The strengths of these matrices are their biodegradability, biocompatibility, and bioactivity (thanks to their mimicry of natural tissues), as well as the versatility of the production process that allows customization for multiple applications.

Previous experiments have already shown their ability to support cell culture while preventing bacterial contamination. Further investigations will assess their antimicrobial performance and ensure no undesirable side effects, leveraging the expertise of the GRAM project research team.

The goal is to develop a product that enhances tissue culture conditions in terms of cell viability, proliferation, and differentiation, enabling simple and reproducible testing for clinical and research applications.

Stakeholders include:

• hospital laboratories (oncological and regenerative medicine applications, pre-natal cell-based testing)
• public and private research laboratories engaged in studies exploiting primary human or animal cells and companies that develop innovate biomaterials and devices for cell culture.