Chan John K, Hamilton Chad A, Anderson Ellen M, Cheung Michael K, Baker Jeanette, Husain Amreen, Teng Nelson N, Kong Christina S, Negrin Robert S
Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of California, San Francisco, UCSF Comprehensive Cancer Center, 94143-1702, USA.
Am J Obstet Gynecol. 2007 Nov;197(5):507.e1-5. doi: 10.1016/j.ajog.2007.05.006.
Primary cancer cells that are extracted from ovarian tumors can serve as an optimal substrate to study the biologic characteristics of ovarian cancer. We describe an efficient and effective method of enriching ovarian tumor cells from ascitic fluid using an immunomagnetic-based method.
Mononuclear cells were isolated from ascites specimens by Ficoll gradient separation. Epithelial ovarian cancer cells were labeled magnetically with monoclonal human epithelial antigen-125 that is conjugated to microbeads. After immunomagnetic separation, the purity of tumor cells before and after purification was quantified by cytologic analysis and confirmed by fluorescence-activated cell sorter analysis.
Peritoneal ascites specimens were obtained from 6 patients with ovarian cancer. The median age of our patients was 61.5 years (range, 46-79 years). Three patients had papillary serous carcinoma; 2 patients had clear cell carcinoma, and 1 patient had an undifferentiated adenocarcinoma. The mean tumor purity was only 22.8% +/- 10% (range, 1%-60%) before separation. After enrichment, the purity improved to 82.3% +/- 4.0% (range, 70%-90%). Our enrichment technique increased the tumor purity by 59.5% +/- 8.4%. The mean percent yield after positive enrichment was 30.1% +/- 14.5%.
The immunomagnetic cell separation technique is an efficient and effective method for isolating and purifying ovarian tumor cells from ascites. Results from experiments with fresh tumor cells rather than cancer cell lines may be more relevant for clinical application.
从卵巢肿瘤中提取的原发性癌细胞可作为研究卵巢癌生物学特性的理想底物。我们描述了一种基于免疫磁珠法从腹水中富集卵巢肿瘤细胞的高效方法。
通过Ficoll梯度分离法从腹水标本中分离出单核细胞。用与微珠偶联的单克隆人上皮抗原-125对上皮性卵巢癌细胞进行磁性标记。免疫磁珠分离后，通过细胞学分析对纯化前后肿瘤细胞的纯度进行定量，并通过荧光激活细胞分选分析进行确认。
从6例卵巢癌患者中获取腹水标本。患者的中位年龄为61.5岁（范围46 - 79岁）。3例为乳头状浆液性癌；2例为透明细胞癌，1例为未分化腺癌。分离前肿瘤细胞的平均纯度仅为22.8%±10%（范围1% - 60%）。富集后，纯度提高到82.3%±4.0%（范围70% - 90%）。我们的富集技术使肿瘤纯度提高了59.5%±8.4%。阳性富集后的平均回收率为30.1%±14.5%。
免疫磁珠细胞分离技术是一种从腹水中分离和纯化卵巢肿瘤细胞的高效方法。使用新鲜肿瘤细胞而非癌细胞系进行实验的结果可能对临床应用更具相关性。
