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2019, Number 4

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Rev Cubana Hematol Inmunol Hemoter 2019; 35 (4)

Design of a multicolored flow cytometry panel for blood, ascites and ovarian tissue samples

Villegas VCA, Torres LG, Morejón MA, Arango PMC

Language: Spanish
References: 29
Page: 1-19
PDF size: 416.11 Kb.


ABSTRACT

Introduction: Epithelial ovarian cancer occupies the 6th place in incidence and mortality in women worldwide. In Cuba, it occupies the 5th place in incidence in females. This cancer is immunogenic and its malignant cells grow in interaction with multiple cells from immune system. Its clinical course depends largely on the type of inflammatory infiltrate accompanying the tumor. Cytology and histopathology are gold standard as diagnostic methods. However, flow cytometry emerges as a technology with greater sensitivity, objectivity and speed. Objective: To design a multicolored flow cytometry panel to immunophenotype the lymphocytic infiltrate of three types of samples for patients with ovarian cancer. Methods: An experimental design was carried out in vitro for the creation and evaluation of a multicolored flow cytometry panel in the Immunology laboratory of the National Institute of Oncology and Radiobiology of Cuba. The panel was designed in the blood of three healthy subjects; then it was optimized for blood in 33 healthy volunteers and blood, ascites and ovarian tumor tissue, from three patients with epithelial ovarian cancer. Several lymphocytes lineages were immunophenotypedin each sample. Results: Eleven markers were selected for the immunophenotype and the panel was made up of four multiparameter cytometry tubes. The methodology created could be applied to the samples of ascites and tumor tissue without interferences and percentages of different lymphocyte subpopulations were obtained within the expected values. Conclusions: The designed panel allowed immunophenotyping of lymphocytes in different types of ovarian cancer patient samples and reliable and reproducible results were obtained. This methodology could be employed for others diseases.


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