Abstract
Introduction. Personalized treatment strategies for embryonal carcinoma, a germ cell tumor, are crucial due to the inherent heterogeneity of the tumor. Existing in vitro models inadequately reflect the native tumor microenvironment. Ex vivo organ culture (EVOC) offers a potential solution by preserving the original tumor architecture and cellular interactions. This study presents the first-ever adaptation of the EVOC platform specifically for non-metastatic embryonal carcinoma, to create a novel tool for studying tumor biology and assessing treatment response.
Purpose of the study. Adaptation of the EVOC platform for non-metastatic embryonal carcinoma to determine the limiting lifespans and develop a universal methodology for assessing cell viability.
Material and methods. Tumor samples were obtained from 23 patients with non-metastatic embryonal carcinoma (cT1 – 3, cN0, M0) who underwent radical orchiectomy. Tissue fragments were cultured in DMEM/F12 medium supplemented with fetal bovine serum and antibiotics. Histological and immunohistochemical analyses were performed on days 0, 3, 7, and 10. The proliferation index (PI) was evaluated using Ki-67 staining. The overall cell number (OCN) and the tumor cell number (TCN; OCT3/4-positive cells) were calculated. To assess tumor viability, the area under the curve (AUC) for the PI was calculated. Statistical analysis included repeated-measures analysis of variance (ANOVA) and post-hoc tests.
Results. Histological examination confirmed the typical morphology of embryonal carcinoma with the preservation of the OCT3/4-positive phenotype throughout the entire observation period. A significant (p < 0.05) decrease in the overall cell number (OCN) and the tumor cell number (TCN) was established by the 10th day of cultivation by 79.9% and 80.6%, respectively. The faster rate of decline in TCN relative to OCN indicates a higher survival capability of the microenvironment components compared to the tumor cells. A consistent decrease in cell proliferative activity (Ki-67) was revealed on days 3, 7, and 10 (p < 0.05). Based on the dynamics of the proliferation index (AUC), a computational method, the “EVOC score”, was proposed as a representative marker of tumor viability for evaluating the effectiveness of various treatment modalities.
Conclusion. We have successfully adapted the EVOC platform for non-metastatic embryonal carcinoma, demonstrating the preservation of viability for up to 10 days. The PI-AUC indicator was determined to be a reliable marker of cell viability.
Keywords
Authors and affiliations
https://elibrary.ru/uavcxp
G.A. Demyashkin(1, 2), D.V. Belokopytov(1), V.I. Shchekin(1), T.G. Borovaya(1), V.V. Rostovskaya(2),
D.A. Tsomartova(2), N.V. Pyatigorskaya(2), O.V. Filippova(2), D.D. Maksarova(1), A.V. Mirontsev(2),
1-P.A. Herzen Moscow Oncology Research Institute – Branch of the National Medical Research Radiological Centre of the Ministry
of Health of the Russian Federation, 2nd Botkinsky proezd, 3, Moscow, 125284, Russian Federation; 2-I.M. Sechenov First
Moscow State Medical University (Sechenov University), Trubetskaya St., 8/2, Moscow, 119048, Russian Federation
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