缓冲液基细胞培养液中储存细胞的细胞形态和生物标志物免疫反应的时间相关性变化。

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

机构信息

Faculty of Medicine, Institute of Pathology, University of Ljubljana, Ljubljana, Slovenia.

Lek Pharmaceuticals d.d., Sandoz Development Center, Ljubljana, Slovenia.

出版信息

Cytopathology. 2021 Jul;32(4):513-518. doi: 10.1111/cyt.12980. Epub 2021 Apr 16.

DOI:10.1111/cyt.12980

PMID:33861498

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.

摘要

目的

缓冲基细胞培养基（BBCM）是细胞学样本采集后处理的一种有价值的工具，尽管其对细胞特性的影响尚未明确。在这项研究中，我们评估了在含有牛血清白蛋白（BSA）和乙二胺四乙酸（EDTA）的 BBCM 中，室温下储存的细胞在时间相关的细胞形态和生物标志物免疫反应性的变化。

方法

从悬浮和储存在 BBCM 中的三种人类细胞系（MCF7、SK-MEL-28、FaDu）中，连续五个 24 小时的时间间隔（0、24、48、72、96）制备细胞离心涂片。通过对巴氏染色的细胞离心涂片上凋亡细胞的百分比来评估细胞形态的保存情况。通过对相应免疫细胞化学反应呈阳性的细胞的百分比来评估细胞角蛋白、雌激素受体、Ki67 和黑色素瘤标志物的免疫反应性的保存情况。

结果

在储存 24 和 48 小时的情况下，三种细胞系的大多数细胞的形态保存良好（分别为 93%、97%、98%和 62%、81%、88%），而大多数细胞在储存 72 和 96 小时后发生凋亡（分别为 70%、47%、39%和 77%、70%、59%）。细胞角蛋白的免疫反应性在整个 96 小时内保持不变，而黑色素瘤标志物（S100、HMB45、Melan-A）的免疫反应性分别下降了 27%、2%和 3%。雌激素受体和 Ki67 的免疫反应性在最初的 24 小时后下降了 29%和 17%，在 96 小时后完全丧失。