Sensitive and Specific Detection of Ewing Sarcoma Minimal Residual Disease in Ovarian and Testicular Tissues in an In Vitro Model
Abstract
:1. Introduction
2. Results
2.1. Experimental Model of EWS Contamination of Germinal Tissues
2.2. Yield of RNA Extraction from Germinal Tissues Frozen Using Slow or Snap Freezing
2.3. Detection of EWS-FLI1 Transcript in Frozen OT and TT Samples Contaminated with RD-ES Cells
2.4. Measurements of Dissemination Potential of RD-ES Cell Lines After Co-Culture with Ovarian and Testicular Samples
2.5. MRD Analysis in Ovarian and Testicular Tissues of Patients with EWS
3. Discussion
4. Materials and Methods
4.1. Patients and Samples
4.2. Freezing and Thawing of Germinal Tissue Samples
4.3. Culture of RD-ES Subclones and Contamination
4.4. Detection of EWS-FLI1 of Type II mRNA Expression by RT-qPCR
4.5. Detection of Potential Dissemination of RD-ES Cells in OT and TT After co-Cultures
4.5.1. OT Co-Culture
Histologic Analysis
Fluorescence in Situ Hybridization (FISH) after Co-Cultures of Ovarian Tissue and RD-ES cells
4.5.2. TT Co-Culture
Immunohistochemistry (IHC)
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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EWS Patients | Age at Cryopreservation (Years) | Chemotherapy | Metastasis | Outcome |
---|---|---|---|---|
OT A | 16 | Before preservation | Lung | Dead |
OT B | 14 | Before preservation | Lung | Alive |
OT C | 14 | Before preservation | No | Alive |
OT D | 15 | Before preservation | Lung, bone and medullary | Dead |
OT E | 13 | Before preservation | Lung | Alive |
TT A | 7 | Before preservation | Mediastinum | Alive |
TT B | 14 | Before preservation | None | Alive |
TT C | 14 | After preservation | None | Alive |
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Chaput, L.; Grèze, V.; Halle, P.; Radosevic-Robin, N.; Pereira, B.; Véronèse, L.; Lejeune, H.; Durand, P.; Martin, G.; Sanfilippo, S.; et al. Sensitive and Specific Detection of Ewing Sarcoma Minimal Residual Disease in Ovarian and Testicular Tissues in an In Vitro Model. Cancers 2019, 11, 1807. https://doi.org/10.3390/cancers11111807
Chaput L, Grèze V, Halle P, Radosevic-Robin N, Pereira B, Véronèse L, Lejeune H, Durand P, Martin G, Sanfilippo S, et al. Sensitive and Specific Detection of Ewing Sarcoma Minimal Residual Disease in Ovarian and Testicular Tissues in an In Vitro Model. Cancers. 2019; 11(11):1807. https://doi.org/10.3390/cancers11111807
Chicago/Turabian StyleChaput, Laure, Victoria Grèze, Pascale Halle, Nina Radosevic-Robin, Bruno Pereira, Lauren Véronèse, Hervé Lejeune, Philippe Durand, Guillaume Martin, Sandra Sanfilippo, and et al. 2019. "Sensitive and Specific Detection of Ewing Sarcoma Minimal Residual Disease in Ovarian and Testicular Tissues in an In Vitro Model" Cancers 11, no. 11: 1807. https://doi.org/10.3390/cancers11111807