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

Area of Science:

• Cytopathology
• Biomarker analysis
• Cell biology

Background:

• Buffer-based cell media (BBCM) are utilized in post-collection cytology sample processing.
• The effects of BBCM on cell morphology and biomarker integrity are not well-defined.
• This study investigates the impact of storage time in BBCM on cell properties.

Purpose of the Study:

• To evaluate time-dependent changes in cell morphology and biomarker immunoreactivity.
• To assess the efficacy of a specific BBCM formulation containing BSA and EDTA.
• To determine optimal storage durations for cytology samples in BBCM at room temperature.

Main Methods:

• Three human cell lines (MCF7, SK-MEL-28, FaDu) were suspended in BBCM with BSA and EDTA.
• Cytospins were prepared at 24-hour intervals up to 96 hours.
• Cell morphology was assessed via Papanicolaou staining (evaluating apoptosis), and biomarker immunoreactivity (cytokeratins, oestrogen receptors, Ki67, melanoma markers) was quantified.

Main Results:

• Cell morphology was well-preserved for up to 48 hours (93-98% intact cells).
• Significant apoptosis occurred after 72 and 96 hours (39-77% apoptotic cells).
• Cytokeratin immunoreactivity remained stable; oestrogen receptors and Ki67 decreased significantly by 24 hours and were lost by 96 hours; melanoma markers showed moderate to minimal loss.

Conclusions:

• BBCM with BSA and EDTA provides good preservation of cell morphology and key biomarker immunoreactivity for up to 48 hours at room temperature.
• Extended storage beyond 48 hours leads to significant cellular degradation and loss of biomarker signals.
• This formulation is suitable for short-term storage of cytology samples, optimizing diagnostic accuracy.