Journal of Cytology
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ORIGINAL ARTICLE
Year : 2022  |  Volume : 39  |  Issue : 2  |  Page : 66-71

Impact of LBC fixative type and fixation time on molecular analysis of pancreatic cancer cells: A comparative study of cell morphology, antigenicity and nucleic acids


1 Graduate School of Health Science, Suzuka University of Medical Science, Kishioka, Suzuka, Mie, Japan
2 Graduate School of Health Science, Suzuka University of Medical Science, Kishioka, Suzuka, Mie, Japan; Clinical Nutrition, Suzuka University of Medical Science, Kishioka, Suzuka, Mie, Japan

Correspondence Address:
Dr. Kazuki Kanayama
Graduate School of Health Science, Suzuka University of Medical Science, 1001-1 Kishioka, Suzuka, Mie 510-0293; Department of Clinical Nutrition, Suzuka University of Medical Science, 1001-1 Kishioka, Suzuka, Mie 510-0293
Japan
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/joc.joc_13_22

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Introduction: Liquid-based cytology (LBC) has been widely used since 2000. Next-Generation Sequencing (NGS) analysis of residual specimens in LBC fixative may also be performed for pancreatic cancer in the near future. We examined cell morphology, antigenicity and nucleic acids in pancreatic cancer cells at different fixation times using two types of LBC fixatives. Methods: PANC-1 cells were fixed in 1 ml CytoRich Red (CR), CytoRich Blue (CB), 95% ethanol (95% AL) or 10% neutral buffered formalin (10% NBF) and evaluated for cell area, antigenicity and nucleic acids with fixation times of 1 hour and 1, 3, 9, and 14 days. Antigenicity was evaluated by immunocytochemical staining for p53 and CK20, and nucleic acid fragmentation was assessed by real-time PCR. Results: There was no difference in total cell area between 1 hour and 14 day fixation times for the CR group, but the CB group showed cell contraction with 9 days fixation. In immunocytochemical staining, the CR group showed high p53 and CK20 positivity even after 14 days fixation. The CB group had a lower p53 positive rate than the CR group from 1 hour fixation. For nucleic acid fragmentation, Ct values for the CR group increased with fixation time. The CB group had consistently low Ct values. Conclusion: Different LBC fixatives and fixation time can have varying effects on cell morphology, antigenicity and nucleic acids in pancreatic cancer cells. Therefore, fixative type and fixation time should be considered for molecular testing on residual samples in LBC fixatives.


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