Isolation of Pancreatic Cancer Cells from a Patient-Derived Xenograft Model Allows for Practical Expansion and Preserved Heterogeneity in Culture
Isolation of Pancreatic Cancer Cells from a Patient-Derived Xenograft Model Allows for Practical Expansion and Preserved Heterogeneity in Culture
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DOI：
10.1016/j.ajpath.2016.02.009
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发表时间：
2016-06-01
影响因子：
6
通讯作者：
Trevino, Jose G.
中科院分区：
文献类型：
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作者：
Pham, Kien;Delitto, Daniel;Trevino, Jose G.
市售的、高度传代的胰腺癌（PC）细胞系的转化价值有限。克服这一限制的尝试主要包括直接从人类 PC 样本中分离和培养癌细胞。然而，这些技术的成功率极低。在这里，我们展示了来自患者来源的异种移植物的原代 PC 细胞系 (PPCL) 的高度可重复培养，其部分保留了已知存在于 PC 中的瘤内异质性。从患者来源的异种移植物中进行 PPCL 扩增的尝试 100% 成功（5 次中的 5 次）。通过流式细胞术、免疫荧光显微镜和短串联重复分析评估表型分析。重要的是，通过皮下注射到非肥胖糖尿病 Cg-Prkdc(scid)Il2rg(tm1Wjl)/SzJ 小鼠中来评估从患者来源的异种移植物扩增的 PPCL 的致瘤性。在形态学上，将所有 PPCL 皮下注射到小鼠体内产生的肿瘤具有与亲本异种移植物相似的特征。 PPCL 一致表达 I 类人类白细胞抗原、上皮细胞粘附分子和细胞角蛋白-19。每个 PPCL 内的细胞表达癌症干细胞标记物 CD44、CD133 和 c-Met 以及免疫标记物人类白细胞抗原 II 类和程序性死亡配体 1 的频率在培养中持续存在异质性。因此，这项工作为快速扩增原代人类 PC 细胞提供了可靠的方法，从而为转化研究提供了一个平台，重要的是，为潜在的个性化治疗方法提供了平台。
Commercially available, highly passaged pancreatic cancer (PC) cell lines are of limited translational value. Attempts to overcome this limitation have primarily consisted of cancer cell isolation and culture directly from human PC specimens. However, these techniques are associated with exceedingly low success rates. Here, we demonstrate a highly reproducible culture of primary PC cell lines (PPCLs) from patient-derived xenografts, which preserve, in part, the intratumoral heterogeneity known to exist in PC. PPCL expansion from patient-derived xenografts was successful in 100% of attempts (5 of 5). Phenotypic analysis was evaluated with flow cytometry, immunofluorescence microscopy, and short tandem repeat profiling. Importantly, tumorigenicity of PPCLs expanded from patient-derived xenografts was assessed by subcutaneous injection into nonobese diabeteic.Cg-Prkdc(scid)Il2rg(tm1Wjl)/SzJ mice. Morphologically, subcutaneous injection of all PPCLs into mice yielded tumors with similar characteristics to the parent xenograft. PPCLs uniformly expressed class I human leukocyte antigen, epithelial cell adhesion molecule, and cytokeratin-19. Heterogeneity within each PPCL persisted in culture for the frequency of cells expressing the cancer stem cell markers CD44, CD133, and c-Met and the immunologic markers human leukocyte antigen class II and programmed death ligand 1. This work therefore presents a reliable method for the rapid expansion of primary human PC cells and, thereby, provides a platform for translational investigation and, importantly, potential personalized therapeutic approaches.