Enrichment of prostate cancer stem cells from primary prostate cancer cultures of biopsy samples

	Enrichment of prostate cancer stem cells from primary prostate cancer cultures of biopsy samples
	Wang, SQ; Huang, SS; Zhao, X; Zhang, QM; Wu, M; Sun, F; Han, G; Wu, DL
刊名	INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL PATHOLOGY
	2014
卷号	7 期号:1 页码:184-193
关键词	Prostate cancer (PrCa) primary prostate cancer cultures (PPrCC) prostate cancer stem cells (PrCSC) sphere culture
通讯作者	Wu, DL (reprint author), Tongji Univ, Dept Urol, Affiliated Tongji Hosp, 389 Xincun Rd, Shanghai 200065, Shanghai, Peoples R China.,ghan@sibs.ac.cn ; wuyy163@126.com
英文摘要	This study was to enrich prostate cancer stem cells (PrCSC) from primary prostate cancer cultures (PPrCC). Primary prostate cancer cells were amplified in keratinocyte serum-free medium with epidermal growth factor (EGF) and bovine pituitary extract (BPE), supplemented with leukemia inhibitory factor (LIF), stem cell factor (SCF) and cholera toxin. After amplification, cells were transferred into ultra-low attachment dishes with serum-free DMEM/F12 medium, supplemented with EGF, basic fibroblast growth factor (bFGF), bovine serum albumin (BSA), insulin, and N2 nutrition. Expression of cell-type-specific markers was determined by RT-qPCR and immunostaining. Tumorigenicity of enriched PrCSC was determined by soft agar assay and xenograft assay in NOD/SCID mice. Biopsy samples from 19 confirmed prostate cancer patients were used for establishing PPrCC, and 18 cases (95%) succeeded. Both basal marker (CK5) and luminal markers (androgen receptor and CK8) strongly co-expressed in most of PPrCC, indicating their basal epithelial origin. After amplification under adherent culture condition in vitro, transient amplifying cells were the dominant cells. Sphere formation efficiency (SFE) of passaged PPrCC was about 0.5%, which was 27 times lower than SFE of LNCaP (13.67%) in the same condition. Compared with adherent cells from PPrCC, prostasphere from PPrCC showed up regulated stem cell markers and increased tumorigenic potential in soft-agar assay. However, spheroid cells from PPrCC prostasphere failed to initiate tumor in xenograft assay in 6 months. Thus, PPrCC can be established and amplified from prostate cancer biopsy samples. Our modified sphere culture system can enrich PrCSC from PPrCC.
学科主题	Oncology; Pathology
类目[WOS]	Oncology ; Pathology
关键词[WOS]	PROSPECTIVE IDENTIFICATION ; PROGENITOR/STEM CELLS ; FORM
收录类别	SCI
语种	英语
WOS记录号	WOS:000335226300018
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/128]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Wang, SQ,Huang, SS,Zhao, X,et al. Enrichment of prostate cancer stem cells from primary prostate cancer cultures of biopsy samples[J]. INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL PATHOLOGY,2014,7(1):184-193.
APA	Wang, SQ.,Huang, SS.,Zhao, X.,Zhang, QM.,Wu, M.,...&Wu, DL.(2014).Enrichment of prostate cancer stem cells from primary prostate cancer cultures of biopsy samples.INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL PATHOLOGY,7(1),184-193.
MLA	Wang, SQ,et al."Enrichment of prostate cancer stem cells from primary prostate cancer cultures of biopsy samples".INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL PATHOLOGY 7.1(2014):184-193.