Zhu Yong-Tong, Wang Chun-Yan, Pang Shi-Yu, Lei Cheng-Yong, Luo Yang, Tan Wan-Long
Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital; Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Department of Neurology, TCM-Integrated Cancer Center of Southern Medical University, Guangzhou, China.
J Cancer Res Ther. 2018 Jun;14(Supplement):S421-S426. doi: 10.4103/0973-1482.174533.
In this study, we showed a modified method for the isolation of cancer stem cells (CSCs) using a combination of differential adhesion method and serum-free culture medium (SFM) method.
Trypsin-sensitive cells and trypsin-resistant cells were isolated from MB49, EJ, and SK-OV-3 cells using a combination of differential adhesion method and SFM method. The CSCs markers expression of trypsin-resistant cells was verified by the flow cytometry, the Western blotting, and the quantitative polymerase chain reaction. Functional comparisons were verified by the resistance to chemotherapy assay, the transwell assay, and the tumor xenograft formation assay.
Trypsin-resistant cells were isolated successfully. They were identified with high expression of CSCs markers and possessed higher resistance to chemotherapy, greater migration in vitro and stronger tumorigenic abilities in vivo.
Trypsin-resistant cells showed specific CSCs characterizations. They were able to be isolated successfully with a modified method by a combination of differential adhesion method and SFM method.
在本研究中，我们展示了一种改良方法，该方法通过结合差异黏附法和无血清培养基（SFM）法来分离癌症干细胞（CSCs）。
使用差异黏附法和SFM法相结合的方式，从MB49、EJ和SK-OV-3细胞中分离出对胰蛋白酶敏感的细胞和对胰蛋白酶耐受的细胞。通过流式细胞术、蛋白质免疫印迹法和定量聚合酶链反应来验证对胰蛋白酶耐受细胞的CSCs标志物表达情况。通过化疗抗性试验、Transwell试验和肿瘤异种移植形成试验来验证功能差异。
成功分离出对胰蛋白酶耐受的细胞。它们被鉴定为具有高表达的CSCs标志物，并且对化疗具有更高的抗性，在体外具有更强的迁移能力，在体内具有更强的致瘤能力。
对胰蛋白酶耐受的细胞表现出特定的CSCs特征。通过差异黏附法和SFM法相结合的改良方法能够成功分离出这些细胞。
