A method for dissociation of oral cancer cell spheroids generated by magnetic 3D bioprinting

Author(s)

Fernandes, Natalie Aparecida Rodrigues ; Nascimento, Camyla Rodrigues ; Coletta, Ricardo D. ; Rossa Junior, Carlos

• Document type: Journal article
• Source: Cytotechnology; v. 77, n. 3, p. 11-pg., 2025-06-01.

Abstract

Spheroid 3D cultures yield a better representation of diverse cellular events in comparison with 2D cultures. However, the ability to investigate some specific phenotype/cell biology-related outcomes of experimental interventions by flow cytometry is somewhat limited in spheroid cultures, as it requires a single cell suspension with good preservation of membrane-bound proteins. In this study, we describe a method combining enzymatic and mechanical dissociation of large spheroids of five different oral squamous cell carcinoma (OSCC) cell lines (H314, SCC9, SCC25, UM-SCC1 and HSC3). Spheroids were generated using magnetic 3D bioprinting. Efficacy of dissociation and viability after dissociation were evaluated by flow cytometry. All spheroids were successfully dissociated into single cells with low cell aggregation using this method, except for spheroids of HSC3 cells. H314 and SCC9 spheroids yielded the highest number of events per microliter and therefore also the largest total number of events. H314 and SCC25 spheroids presented the better results for cell viability after dissociation. H314 spheroids also gave the largest total number of viable cell events. The dissociation method described can be useful for researchers interested in flow cytometry-based analysis of spheroid cultures. (AU)

• FAPESP's process: 20/10664-7 - Contribution of the tissue microenvironment to OSCC progression: role of CAFs, TAMs and fibronectin / a5b1 integrin signaling
• Grantee: Carlos Rossa Junior
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/10544-1 - Contribution of the tissue microenvironment to OSCC progression: role of CAFs, TAMs and fibronectin/a5b1 integrin signaling
• Grantee: Natalie Aparecida Rodrigues Fernandes
• Support Opportunities: Scholarships in Brazil - Doctorate