Authentication of newly established human esophageal squamous cell carcinoma cell line (YM-1) using short tandem repeat (STR) profiling method
Authentication of newly established human esophageal squamous cell carcinoma cell line (YM-1) using short tandem repeat (STR) profiling method
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DOI：
10.1007/s13277-015-4133-4
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发表时间：
2016-03-01
期刊：
影响因子：
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通讯作者：
Jahanbakhsh, Asadi
中科院分区：
文献类型：
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作者：
Ayyoob, Khosravi;Masoud, Khoshnia;Jahanbakhsh, Asadi
在新细胞系建立期间或建立后早期的交叉污染可能导致错误识别的细胞系在世界范围内传播。因此，新建立的细胞系需要用参考标准方法进行鉴定。本研究采用短串联重复序列(STR)DNA图谱技术，对新建立的人食管鳞癌(ESCC)上皮细胞系YM-1的真实性进行了研究。原代培养的人ESCC上皮细胞取自1例成年女性患者的新鲜肿瘤组织。对YM-1细胞的生长特性和上皮来源进行了研究。从第22代收获的YM-1细胞和两个不同日期的ESCC供体肿瘤样本中提取基因组DNA，以防止可能的DNA污染。用AmpFa“”Stra(R)IdentifilerA(R)Plus聚合酶链式反应扩增试剂盒进行STR分析。为了解决YM-1细胞是否会随着传代次数的增加而发生基因改变，在第51代时再次对收获的细胞进行STR图谱分析。YM-1细胞呈单层生长，群体倍增时间为40.66 h，细胞角蛋白AE1/AE3的ICC/IF染色证实YM-1细胞具有上皮源性。在第22代时，ESCC供体肿瘤样本的STR图谱与YM-1细胞相同。然而，供体肿瘤样本的STR图谱显示，他们的D7S820基因座上存在一个非阶梯(OL)等位基因。此外，51代YM-1细胞的重组显示D18S51基因座的杂合性丢失(LOH)。这表明，长期培养细胞系可能会改变它们的DNA图谱。比较DSMZ和ATCC STR图谱数据库中的DNA指纹图谱结果，证实了YM-1细胞系的独特身份。本研究为新建立的细胞系的鉴定提供了一种简便、快速、可靠的方法，有助于防止误认细胞的扩散，从而提高实验的重复性和有效性。
Cross-contamination during or early after establishment of a new cell line could result in the worldwide spread of a misidentified cell line. Therefore, newly established cell lines need to be authenticated by a reference standard method. This study was conducted to investigate the authenticity of a newly established epithelial cell line of human esophageal squamous cell carcinoma (ESCC) called YM-1 using short tandem repeat (STR) DNA profiling method. Primary human ESCC epithelial cells were cultured from the fresh tumor tissue of an adult female patient. Growth characteristics and epithelial originality of YM-1 cells were studied. Genomic DNA was isolated from YM-1 cells harvested at passage 22 and ESCC donor tumor sample on two different days to prevent probable DNA contamination. STR profiling was performed using AmpFa""STRA (R) IdentifilerA (R) Plus PCR Amplification Kit. To address whether YM-1 cells undergo genetic alteration as the passage number increases, STR profiling was performed again on harvested cells at passage 51. YM-1 cells grew as a monolayer with a population doubling time of 40.66 h. Epithelial originality of YM-1 cells was confirmed using ICC/IF staining of cytokeratins AE1/AE3. The STR profile of the ESCC donor tumor sample was the same with YM-1 cells at passage 22. However, STR profile of the donor tumor sample showed an off-ladder (OL) allele in their D7S820 locus. Also, re-profiling of YM-1 cells at passage 51 showed a loss of heterozygosity (LOH) at D18S51 locus. This suggests that long-term culture of cell lines may alter their DNA profile. Comparison of the DNA fingerprinting results in DSMZ, and ATCC STR profiling databases confirmed unique identity of YM-1 cell line. This study provides an easy, fast, and reliable procedure for authentication of newly established cell lines, which helps in preventing the spread of misidentified cells and improving the reproducibility and validity of experiments, consequently.