Tricoli Lucas, Naeem Aisha, Parasido Erika, Mikhaiel John P, Choudhry Muhammad Umer, Berry Deborah L, Abdelgawad Iman A, Lee Richard J, Feldman Adam S, Ihemelandu Chukwuemeka, Avantaggiati Maria, Kumar Deepak, Byers Stephen, Gallagher Rosa, Wulfkuhle Julia, Petricoin Emanuel, Rodriguez Olga, Albanese Chris
Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC, USA.
Julius L. Chambers Biomedical/Biotechnology Research Institute, North Carolina Central University, Durham, NC, USA.
Oncotarget. 2017 Dec 18;9(2):2193-2207. doi: 10.18632/oncotarget.23363. eCollection 2018 Jan 5.
The inability to propagate human prostate epithelial cells indefinitely has historically presented a serious impediment to prostate cancer research. The conditionally reprogrammed cell (CRC) approach uses the combination of irradiated J2 mouse fibroblasts and a Rho kinase inhibitor such as Y27632 to support the continuous culture of cells derived from most epithelial tissues, including the prostate. Due to their rapid establishment and overall ease of use, CRCs are now widely used in a variety of basic and preclinical settings. In addition, CRCs were successfully used to clinically treat respiratory papillomatosis. Although both normal and tumor-derived prostate CRCs have been used to study the basic biology of prostate cancer and to test new therapies, certain limitations exist. We have previously reported that prostate CRCs form functional prostate glands when implanted under the mouse renal capsule. However in conventional culture, the prostate CRCs exist in an adult stem-like, transient amplifying state and consequently do not adequately recapitulate several important features of a differentiated prostate epithelium. To address these limitations, we previously described a transwell dish-based model that supported the culturing of prostate CRCs and the collection of cells and cell extracts for molecular and genetic analyses. Using normal and tumor-derived prostate CRCs, we describe the combined effects of the multi-dimensional transwell platform and defined culture media on prostate cellular proliferation, differentiation and signaling.
长期以来，无法无限传代培养人前列腺上皮细胞一直是前列腺癌研究的严重障碍。条件重编程细胞（CRC）方法利用经辐照的J2小鼠成纤维细胞和一种Rho激酶抑制剂（如Y27632）的组合，来支持源自包括前列腺在内的大多数上皮组织的细胞的连续培养。由于其建立迅速且总体易于使用，CRCs现在广泛应用于各种基础和临床前研究环境。此外，CRCs已成功用于临床治疗呼吸道乳头状瘤病。尽管正常和肿瘤来源的前列腺CRCs都已用于研究前列腺癌的基础生物学和测试新疗法，但仍存在某些局限性。我们之前报道过，前列腺CRCs植入小鼠肾被膜下时会形成功能性前列腺腺体。然而在传统培养中，前列腺CRCs处于成体干细胞样的短暂扩增状态，因此不能充分重现分化前列腺上皮的几个重要特征。为了解决这些局限性，我们之前描述了一种基于Transwell培养皿的模型，该模型支持前列腺CRCs的培养以及用于分子和遗传分析的细胞及细胞提取物的收集。利用正常和肿瘤来源的前列腺CRCs，我们描述了多维Transwell平台和特定培养基对前列腺细胞增殖、分化和信号传导的联合作用。
