Researchers developed a simplified method for creating patient-derived skin tumor organoids that preserved immune cells and accurately reflected clinical tissue characteristics, according to a study published in Cell Organoid. The technique, which required neither enzymatic dissociation nor Matrigel support, produced organoids within 30-60 minutes.
The study demonstrated that dermatofibrosarcoma protuberans (DFSP) organoids maintained up to 11 distinct cell types and exhibited genetic profiles matching their source tissues with approximately 80% concordance in mutation sites. However, there was less than 40% overlap in mutation sites between patients, indicating significant genetic heterogeneity among them.
When tested with imatinib (10 μmol/L) and metformin (2.5 mmol/L), the organoids showed measurable responses. Metformin modulated immune signaling pathways and demonstrated efficacy from day 6 to day 13 of treatment.
The research included samples from 10 skin tumor patients: 6 with DFSP, 2 with keloid, 1 with melanoma, and 1 with malignant schwannoma. Single-cell sequencing identified various cell types within the organoids, including endothelial cells, fibroblasts, keratinocytes, melanocytes, and immune cells such as T cells and mast cells. DFSP organoids consistently exhibited PRSS3 gene mutation patterns within matched tissue-organoid pairs.
The organoids demonstrated distinct morphological patterns: SK75 exhibited spindle-shaped cancer cells, SK76 showed multiple adipocytes, and SK77 displayed disordered cells. All reached volumes of 2 mm³, though growth speeds varied among different generations without correlation to passage number.
Metformin treatment resulted in upregulation of cell cycle, phagocytosis, B cell activation, and P53 signaling pathways, while downregulating DNA transcription, apoptotic pathway, innate-immune response, and Wnt signaling. SK74 and SK75 showed significant response to imatinib, while SK76 did not.
The tissue samples were processed using 100-200 μL of culture medium initially, followed by 1 mL for resuspension, and 15 mL for maintenance. Tissue blocks were cut into 0.5-1 mm³ microtissue blocks for culture.
DFSP organoids showed an abundance of fibroblast cells with relatively scarce immune cells, while keloid organoids displayed a higher presence of keratinocytes and T cells, suggesting diverse immune microenvironments in cultured organoids from different diseases.
Declaration of conflicting interests can be found in the study.
