Standardized Upstream Processing of Ovarian Cancer Surgical Specimens for TIL-Oriented Cellular Workflows

Background: Fresh ovarian cancer tissue is a demanding starting material for cellular workflows, particularly when the aim is to obtain tumor-derived suspensions suitable for further TIL-oriented procedures. In this study, we evaluated post-isolation cell concentration and viability in ovarian cancer specimens processed according to a standardized mechanical–enzymatic protocol. Methods: Biological material was collected from 24 patients undergoing routine surgery for ovarian cancer or suspected advanced ovarian malignancy. After macroscopic assessment and exclusion of extensively necrotic tissue, 19 specimens were included in the final analysis. The cohort was dominated by high-grade serous ovarian carcinoma, which accounted for 15 of 19 analyzed cases. Tissue specimens varied markedly in mass, with a median processed tissue weight of 2.05 g and a range from 0.11 to 14.93 g. Results: The median final cell concentration was 8.10 × 106 cells/mL, with values ranging from 6.40 × 103 to 9.26 × 108 cells/mL. Median viability was 80.8%, although individual results ranged widely from 2.3% to 98.7%. The calculated median concentration of viable cells was 6.17 × 106 viable cells/mL. Tissue mass showed a moderate positive association with final cell concentration (Spearman’s rho = 0.50, p = 0.028), but not with viability. Conclusions: These findings show that standardized processing of ovarian cancer surgical specimens can generate cell suspensions suitable for post-isolation quality assessment, even when the input material is highly variable. The present study focused on early post-isolation quality-control readouts obtained after tumor dissociation and did not evaluate TIL expansion capacity, detailed TIL phenotype, or antitumor function. The wide range of final cell concentrations underlines the biological and practical heterogeneity of ovarian cancer tissue and supports the need for careful reporting of early tissue-processing parameters in TIL-oriented workflows.