Construction of primary prostate cancer cell bank using conditional reprogramming cell technology
10.3760/cma.j.cn112330-20191204-00539
- VernacularTitle:利用条件性重编程细胞技术构建前列腺癌原代细胞库
- Author:
Jianli DUAN
1
;
Zifeng WANG
;
Weijiao YANG
;
Hanqi LEI
;
Junfu ZHANG
;
Yonghong LI
;
Jun PANG
Author Information
1. 中山大学附属第三医院泌尿外科，广州 510630
- From:
Chinese Journal of Urology
2020;41(5):374-381
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To cultivate human-derived prostate cancer (PCa) cells via conditional reprogramming cell (CRC) technology, and establish individualized cell bank for PCa research in vitro.Methods:We obtained three fresh PCa tissue samples from different patients between January 2019 and April 2019. Then each sample was divided into two parts. One was used for cancer nature confirmation by intraoperative biopsy. Another part was sent to the laboratory and digested into single primary cancer cells with 0.25% EDTA enzyme for CRC technology. The details were described as followed: 1. The primary PCa cells were co-cultured with 3T3-J2 cells irradiated by 30 Gy (feeder cells) in conditioned medium, and observed for the growth of cell clones, 2. The feeder cells were removed by 0.25% EDTA trypsin for 1 minute before primary PCa cells digested for passage. All primary PCa cells were validated by multiple experiments such as immunofluorescence, immunohistochemistry, immunoblotting and fluorescence in situ hybridization (FISH).Results:Total three cases of human-derived PCa cell lines were successfully established during 15days through CRC technology. All those primary PCa cells could be steadily and continuously passaged, which also expressed AR, CK5, CK18, P504s and PSA. FISH demonstrated that each cell line harbored≥1.6% TMPRSS2/ERG fusion and conformed to the features of PCa.Conclusion:CRC technology can be used for stable and continuous PCa cell culture in vitro.
