Time‐related changes in cell morphology and biomarker immunoreactivity for cells stored in a buffer‐based cell medium

Abstract

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.

Pheochromocytoma presenting as a pancreatic mass: Avoiding a dangerous pitfall in endoscopic ultrasound‐guided fine needle aspiration biopsy using GATA3 immunostain

“…The rest of the sample was stored in a laboratory-made cell medium. 13 ROSE confirmed abundant specimens containing neoplastic cells (Figure 2A). Cytospins and a cell block were prepared in the laboratory.…”

“…Two smears were prepared with each pass—Hemacolor for rapid on‐site evaluation (ROSE), and May‐Grünwald‐Giemsa (MGG) stain. The rest of the sample was stored in a laboratory‐made cell medium 13 . ROSE confirmed abundant specimens containing neoplastic cells (Figure 2A).…”

Role of endoscopic ultrasound-guided fine needle aspiration biopsies in diagnosing pancreatic neoplasms in the paediatric population: experience from a tertiary center and review of the literature

“…The remaining sample was rinsed and stored in an in-house made cell medium. 7 There was a maximum of 5 passes in one case with initial unsatisfactory samples (e.g. samples without diagnostic cells).…”

Optimization of pre‐analytical and analytical steps for DNA and RNA analysis of fresh cytology samples

“…In our laboratory, highly effective standardized processing of cytology samples for different ancillary diagnostic methods was developed. After preparation of direct smears for ROSE and cytomorphological evaluation from fine‐needle aspiration biopsies (free‐hand and image‐guided) or effusions, the sample leftovers are suspended in a buffer‐based cell medium (BBCM) with bovine serum albumin (BSA) and ethylene diamine tetraacetic acid (EDTA) and stored at room temperature 23 …”

“…After preparation of direct smears for ROSE and cytomorphological evaluation from fine-needle aspiration biopsies (free-hand and imageguided) or effusions, the sample leftovers are suspended in a buffer-based cell medium (BBCM) with bovine serum albumin (BSA) and ethylene diamine tetraacetic acid (EDTA) and stored at room temperature. 23 These cell suspensions, prepared from different cytology samples and stored at RT, can be used in a timeframe of up to 72 h for the preparation of additional cytospins for immunocytochemistry, [24][25][26] or special staining's as well as for flow cytometry immunophenotyping 28,29 or even for the preparation of cell blocks. 30 Moreover the suitability of each cell suspension for further processing is evaluated using ROSE on test cytospin.…”