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Time‐related changes in cell morphology and biomarker immunoreactivity for cells stored in a buffer‐based cell medium
Author(s) -
Srebotnik Kirbis Irena,
Prosen Lara,
Strojan Flezar Margareta
Publication year - 2021
Publication title -
cytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 48
eISSN - 1365-2303
pISSN - 0956-5507
DOI - 10.1111/cyt.12980
Subject(s) - cell , receptor , cell culture , andrology , bovine serum albumin , medicine , morphology (biology) , apoptosis , cell counting , cytology , staining , pathology , microbiology and biotechnology , chemistry , biology , immunology , biochemistry , cell cycle , genetics
Objective Buffer‐based cell media (BBCM) are a valuable tool in the post‐collection processing of cytology samples, though with poorly defined effects on cell properties. In this study, time‐related changes in cell morphology and biomarker immunoreactivity were evaluated for cells stored at room temperature in a BBCM prepared with bovine serum albumin (BSA) and ethylene diamine tetraacetic acid (EDTA). Methods Cytospins were prepared at five consecutive 24‐hour intervals (0, 24, 48, 72, 96) from three human cell lines (MCF7, SK‐MEL‐28, FaDu) suspended and stored in BBCM. Preservation of cell morphology was evaluated on Papanicolaou‐stained cytospins from the percentages of apoptotic cells. Preservation of immunoreactivity was evaluated for cytokeratins, oestrogen receptors, Ki67, and melanoma markers from the percentages of cells positive for the corresponding immunocytochemical reactions. Results Cell morphology was well preserved for the majority of cells of the three lines stored for 24 and 48 hours (93%, 97%, 98% and 62%, 81%, 88%, respectively), while the majority of cells were apoptotic after 72 and 96 hours (70%, 47%, 39% and 77%, 70%, 59%, respectively). The immunoreactivity of cytokeratins remained unchanged during the entire 96 hours, while that of melanoma markers (S100, HMB45, Melan‐A) decreased by 27%, 2%, and 3%, respectively. The immunoreactivity of oestrogen receptors and Ki67 decreased by 29% and 17% after the first 24 hours, and was completely lost after 96 hours. Conclusions A BBCM with the addition of BSA and EDTA facilitates good preservation of cell morphology and immunoreactivity of biomarkers for up to 48 hours at room temperature.