Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, Japan.
Division of Biochemistry, Chemistry Department, Faculty of Science, Menoufia University, Shebin El Koum-Menoufia, Egypt.
Methods Mol Biol. 2022;2549:399-407. doi: 10.1007/7651_2021_371.
Cancer stem cells (CSCs) are capable of continuous proliferation and self-renewal and are proposed to play significant roles in oncogenesis, tumor growth, metastasis, and cancer recurrence. CSCs are considered derived from normal stem cells affected by the inflammatory microenvironment. Stem cells, are considered to be induced into progenitor cells, which differentiate into various normal phenotypes depending on the normal niche. We hypothesized that CSCs could be derived from stem cells in the cancer-inducing niche, which is a condition of chronic inflammation rich in growth factors, interleukins, chemokines, etc. Exosomes are considered to be the key mediators responsible for the cell-to-cell communications carrying proteins, nucleic acids, metabolites, etc., to shuttle between cells. If these cells are in the environment of chronic inflammation, the exosomes should be reflecting the conditions. In this chapter, we detail the method of CSC initiation using extracellular vesicles (EVs) derived from cancer cell. The stem cells treated with the EVs acquired characteristics of CSCs showing spheroids expressing stemness markers in the suspension culture and high tumorigenicity in Balb/c nude mice. EVs might perform as suitable inducer for initiating CSCs from stem cells or progenitor cells. The model of CSCs and the procedure of their establishment with EVs will help study the exact effect of EVs in the cancer-inducing niche and tumor microenvironment.
癌症干细胞（CSCs）具有持续增殖和自我更新的能力，被认为在肿瘤发生、肿瘤生长、转移和癌症复发中发挥重要作用。CSCs 被认为来源于受炎症微环境影响的正常干细胞。干细胞被认为是诱导成祖细胞的，根据正常生态位，祖细胞分化为各种正常表型。我们假设 CSCs 可以来源于致癌生态位中的干细胞，这是一种富含生长因子、白细胞介素、趋化因子等的慢性炎症条件。外泌体被认为是负责携带蛋白质、核酸、代谢物等在细胞间进行细胞间通讯的关键介质，将它们在细胞间穿梭。如果这些细胞处于慢性炎症环境中，外泌体应该反映出这种情况。在本章中，我们详细介绍了使用源自癌细胞的细胞外囊泡（EVs）启动 CSC 的方法。用 EVs 处理的干细胞获得了 CSC 的特征，在悬浮培养中表现出表达干细胞标志物的球体，并在 Balb/c 裸鼠中具有高致瘤性。EVs 可能作为合适的诱导剂，从干细胞或祖细胞中诱导 CSC 的产生。CSC 模型及其用 EVs 建立的过程将有助于研究 EVs 在致癌生态位和肿瘤微环境中的确切作用。
