Yaremko M L, Kelemen P R, Kutza C, Barker D, Westbrook C A
Department of Pathology, University of Chicago, Illinois 60637, USA.
Am J Pathol. 1996 Jan;148(1):95-104.
Immunomagnetic separation is a highly specific technique for the enrichment or isolation of cells from a variety of fresh tissues and microorganisms or molecules from suspensions. Because new techniques for molecular analysis of solid tumors are now applicable to fixed tissue but sometimes require or benefit from enrichment for tumor cells, we tested the efficacy of immunomagnetic separation for enriching fixed solid tumors for malignant epithelial cells. We applied it to two different tumors and fixation methods to separate neoplastic from non-neoplastic cells in primary colorectal cancers and metastatic breast cancers, and were able to enrich to a high degree of purity. Immunomagnetic separation was effective in unembedded fixed tissue as well as fixed paraffin-embedded tissue. The magnetically separated cells were amenable to fluorescence in situ hybridization and polymerase chain reaction amplification of their DNA with minimal additional manipulation. The high degree of enrichment achieved before amplification contributed to interpretation of loss of heterozygosity in metastatic breast cancers, and simplified fluorescence in situ hybridization analysis because only neoplastic cells were hybridized and counted. Immunomagnetic separation is effective for the enrichment of fixed solid tumors, can be performed with widely available commercial antibodies, and requires little specialized instrumentation. It can contribute to interpretation of results in situations where enrichment by other methods is difficult or not possible.
免疫磁珠分离是一种高度特异性的技术，可用于从各种新鲜组织、微生物中富集或分离细胞，或从悬浮液中分离分子。由于目前用于实体瘤分子分析的新技术适用于固定组织，但有时需要对肿瘤细胞进行富集或能从富集过程中受益，我们测试了免疫磁珠分离法对固定实体瘤中恶性上皮细胞的富集效果。我们将其应用于两种不同的肿瘤及固定方法，以分离原发性结直肠癌和转移性乳腺癌中的肿瘤细胞与非肿瘤细胞，并能够实现高度纯化。免疫磁珠分离法在未包埋的固定组织以及固定石蜡包埋组织中均有效。经磁珠分离的细胞适用于荧光原位杂交及DNA的聚合酶链反应扩增，且只需进行极少的额外操作。扩增前实现的高度富集有助于对转移性乳腺癌杂合性缺失的解读，并且简化了荧光原位杂交分析，因为仅对肿瘤细胞进行杂交和计数。免疫磁珠分离法对固定实体瘤的富集有效，可使用广泛可得的商业抗体进行操作且所需的专业仪器很少。在其他方法富集困难或无法进行富集的情况下，它有助于结果的解读。
