Research Service, Minneapolis Veterans Affairs Healthcare System, Minneapolis, MN, U.S.A.
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, U.S.A.
Anticancer Res. 2019 Aug;39(8):4171-4177. doi: 10.21873/anticanres.13576.
BACKGROUND/AIM: Identification of prostatic stem cells in primary prostate tissue sections, organ cultures of prostate and cell lines requires a range of techniques that allows characterization of stem cells for their potential use in the treatment of patients. Isolated cells usually round-up and develop changes in shape, size and cellular characteristics. The aim of this study was to provide a range of methods for identifying prostatic stem cells and characterizing them with regard to ultrastructure, nuclear morphology, cytoplasmic organelles, and/or expression stem cell marker CD133.
Prostate biopsy and prostatectomy specimens were used for studying prostatic stem cells and their known marker CD133 in tissue sections by light and/or electron microscopy. Inverted capsule embedding was used to study archival metastatic prostate in pelvic nodes and Du145 cell line in a monolayer culture.
Staining for CD133 positively identified stem cells that were found in benign prostatic hyperplasia, benign prostate, and prostate cancer cells. Paraffin embedded sections showed a single type of stem cells, whereas methylene blue-stained Epon sections showed both light and dark stem cells. Electron microscopy showed that both basal and stem cells were closely associated with the basement membrane (basal lamina). Stem cells had smooth plasma and nuclear membranes, a prominent nucleolus, small mitochondria, and few ribosomes. Du145 cells were separated by intercellular spaces in monolayer culture.
The inverted capsule embedding method allowed the study of metastasized prostate cancer in pelvic lymph nodes. Our approach enabled the assessment of stem cells in tissue sections by light and electron microscopy.
背景/目的：在原发性前列腺组织切片、前列腺器官培养物和细胞系中识别前列腺干细胞需要一系列技术，这些技术可用于鉴定干细胞，以将其潜在应用于患者的治疗。分离的细胞通常会圆化，并在形状、大小和细胞特征方面发生变化。本研究旨在提供一系列鉴定前列腺干细胞的方法，并从超微结构、核形态、细胞质细胞器和/或表达干细胞标志物 CD133 方面对其进行特征描述。
使用前列腺活检和前列腺切除术标本，通过光镜和/或电镜研究组织切片中的前列腺干细胞及其已知标志物 CD133。采用反转胶囊包埋法研究盆腔淋巴结转移性前列腺和单层培养中的 Du145 细胞系。
CD133 染色可阳性鉴定出存在于良性前列腺增生、良性前列腺和前列腺癌细胞中的干细胞。石蜡包埋切片显示单一类型的干细胞，而亚甲蓝染色的 Epon 切片显示亮细胞和暗细胞。电镜显示，基底细胞和干细胞均与基底膜（基底膜）紧密相关。干细胞具有光滑的质膜和核膜、明显的核仁、小的线粒体和较少的核糖体。在单层培养中，Du145 细胞通过细胞间空间分离。
反转胶囊包埋法可用于研究盆腔淋巴结中的转移性前列腺癌。我们的方法使我们能够通过光镜和电镜评估组织切片中的干细胞。
