Журнал исследований и терапии стволовых клеток

Абстрактный

TRA-1-60+, SSEA-4+, Oct4A+, Nanog+ Clones of Pluripotent Stem Cells in the Embryonal Carcinomas of the Ovaries

Marek Malecki, Mark Anderson, Michael Beauchaine, Songwon Seo, Xenia Tombokan and Raf Malecki

Introduction:
Embryonal Carcinoma of the Ovary (ECO), pure or admixed to other tumors, is the deadly gynecological cancer.
Specific aim: The specific aim of this work was identification, isolation, clonal expansion, and molecular profiling of the pluripotent cells in the embryonal carcinomas of the ovaries.
Patients and methods: The samples were collected from the patients, who were clinically and histopathologically diagnosed with the advanced, pure ECO. Preparation of the samples was initiated by negative selection of the cells by MACS, while using the superparamagnetic scFvs against phosphatidylserine (PS), and dsDNA, CD45, CD34, CD19, CD14, and by positive selection, while using the superparamagnetic scFvs for TRA-1-60 and SSEA-4. The cell surface display was analyzed by flow cytometry (FCM), immunoblotting (IB), multiphoton fluorescence spectroscopy (MPFS), nuclear magnetic resonance spectroscopy (NMRS), and total reflection x-ray spectroscopy (TRXFS). The transcripts of the OCT4A and Nanog were analyzed by qRTPCR and MPFS. The human pluripotent, embryonic stem cells (ESC), human pluripotent, embryonal carcinoma of the testes (ECT), healthy tissues of the ovary (HTO), healthy tissue of the testes, peripheral blood mononuclear cells (PBMC), and bone marrow mononuclear cells (BMMC) served as the controls.
Results: The studied embryonal carcinomas of the ovary (ECO) contained the cells with significantly higher intensity of the surface display of the TRA-1-60 and SSEA-4, relative to the BMMC, PBMC, and HTO, but similar to the pluripotent ESC and ECT. Their morphology and ultrastructure were consistent with the histopathological diagnoses. Moreover, these cells were significantly stronger expressers of the Oct4A and Nanog, relative to the PBMC, BMMC, and HTO, but similar relative to the pluripotent ESC and ECT. The ECO cells formed embryoid bodies, which differentiated into ectoderm, mesoderm, and endoderm. These cells were induced to differentiate into muscles, epithelia, and neurons.
Conclusion: Herein, we revealed presence and identify molecular profiles of the clones of the pluripotent stem cells in the embryonal carcinomas of the ovaries. These results should help us with refining molecular diagnoses of these deadly neoplasms.