用于分离和鉴定假定前列腺干细胞的新方法。
Novel method for the isolation and characterisation of the putative prostatic stem cell.
作者信息
Bhatt Rupesh I, Brown Michael D, Hart Claire A, Gilmore Paul, Ramani Vijay A C, George Nicholas J, Clarke Noel W
机构信息
Genito-Urinary Cancer Research Group, Cancer Research UK Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK.
出版信息
Cytometry A. 2003 Aug;54(2):89-99. doi: 10.1002/cyto.a.10058.
BACKGROUND
Prostate stem cells, responsible for the development, maturation, and function of the prostate, have been implicated in the aetiology of both benign prostate hyperplasia (BPH) and prostate cancer (CaP). However, research has been hampered by the lack of a definitive stem cell marker. We have adapted the protocol for differential Hoechst 33342 uptake by hemopoietic stem cells to enable isolation of putative stem cells from the prostate.
METHODS
Prostate epithelial cells isolated from prostate tissue obtained from patients with BPH after transurethral resection of the prostate were stained with Hoechst 33342. The Hoechst 33342 Red/Blue flow cytometry profile was then determined. Hoechst 33342 and Pyronin Y staining was used to determined the cell cycle status.
RESULTS
A verapamil-sensitive side population (SP) can be isolated from primary prostate tissue accounting for 1.38% +/- 0.07% of prostate epithelial cells. Cell cycle analysis of this SP population revealed that the majority of SP cells are in either G0 (12.38 +/- 0.31%) or G1 (63.19 +/- 2.13%).
CONCLUSIONS
The Hoechst 33342 dye efflux protocol can be adapted for the isolation of a SP from primary prostate tissue.
背景
前列腺干细胞负责前列腺的发育、成熟和功能，已被认为与良性前列腺增生（BPH）和前列腺癌（CaP）的病因有关。然而，由于缺乏明确的干细胞标志物，相关研究受到了阻碍。我们采用了造血干细胞对Hoechst 33342摄取差异的方案，以从前列腺中分离出假定的干细胞。
方法
对经尿道前列腺切除术后从BPH患者前列腺组织中分离出的前列腺上皮细胞进行Hoechst 33342染色。然后测定Hoechst 33342红/蓝流式细胞术图谱。使用Hoechst 33342和派洛宁Y染色来确定细胞周期状态。
结果
可以从原发性前列腺组织中分离出对维拉帕米敏感的侧群细胞（SP），其占前列腺上皮细胞的1.38%±0.07%。对该SP群体的细胞周期分析显示，大多数SP细胞处于G0期（12.38±0.31%）或G1期（63.19±2.13%）。
结论
Hoechst 33342染料外排方案可用于从原发性前列腺组织中分离出SP。
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