Eskişehir Osmangazi University, Cellular Therapy and Stem Cell Production Application and Research Center, Eskişehir, Turkey
İstinye University, Faculty of Medicine, Department of Histology and Embryology, İstanbul, Turkey
Turk J Haematol. 2021 Dec 7;38(4):254-263. doi: 10.4274/tjh.galenos.2021.2020.0682. Epub 2021 Mar 24.
Patient-specific induced pluripotent stem cells (iPSCs) have potential in human disease modeling and regenerative medicine. The in vitro phenotype of disease-specific iPSC-derived cells can be used to bridge the knowledge gap between clinical phenotype and molecular or cellular pathophysiology and to understand the pathology of diseases, along with further applications, such as creating new strategies for drug screening or developing novel therapeutic agents. The aim of our study was to generate iPSCs from multiple myeloma (MM) patients.
Mesenchymal stem cells (MSCs) isolated from MM patients were induced for pluripotency via the Sendai virus. Fibroblasts were used as a control. Microscopic analysis was performed daily. For colony selection, live staining was done using alkaline phosphatase staining. Reprogramming experiments were confirmed by flow cytometry, immunofluorescence (IF) staining, and gene expression analyses. To confirm the spontaneous differentiation potential, an in vitro embryonic body (EB) formation assay was performed.
Fibroblasts and MSCs obtained from MM patients were reprogrammed using the Sendai virus, which contains reprogramming vectors with the four Yamanaka factors, Oct3/4, Sox2, Klf4, and c-Myc. Microscopic analysis revealed that the generated iPSCs possessed classical embryonic stem cell-like morphological characteristics. Reprogramming experiments further showed that both cell lines can be reprogrammed up to the pluripotent stage, which was confirmed by flow cytometry, IF staining, and gene expression analyses. Spontaneous differentiation potential was confirmed by in vitro EB formation assays.
iPSCs have been successfully obtained from MM patients for the first time. These cells could clarify the molecular mechanisms behind this disease.
患者特异性诱导多能干细胞（iPSC）在人类疾病建模和再生医学中有很大的应用潜力。疾病特异性 iPSC 衍生细胞的体外表型可用于弥合临床表型与分子或细胞病理生理学之间的知识空白，并帮助理解疾病的病理学，同时还可以进一步应用，例如创建新药筛选的新策略或开发新的治疗药物。本研究旨在从多发性骨髓瘤（MM）患者中生成 iPSC。
通过仙台病毒将从 MM 患者中分离出的间充质干细胞（MSC）诱导为多能性细胞。纤维母细胞被用作对照。每日进行显微镜分析。对于集落选择，使用碱性磷酸酶染色进行活细胞染色。通过流式细胞术、免疫荧光（IF）染色和基因表达分析确认重编程实验。为了确认自发分化潜能，进行了体外胚胎体（EB）形成测定。
使用包含四个山中因子（Oct3/4、Sox2、Klf4 和 c-Myc）的重编程载体的仙台病毒，对来自 MM 患者的纤维母细胞和 MSC 进行了重编程。显微镜分析显示，生成的 iPSC 具有典型的胚胎干细胞样形态特征。重编程实验进一步表明，这两种细胞系都可以被重编程至多能性阶段，这通过流式细胞术、IF 染色和基因表达分析得到了证实。通过体外 EB 形成测定证实了自发分化潜能。
首次从 MM 患者中成功获得了 iPSC。这些细胞可以阐明该疾病背后的分子机制。
