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评估源自输卵管和皮肤起源的诱导多能干细胞的输卵管上皮细胞特征。

Assessment of Fallopian Tube Epithelium Features Derived from Induced Pluripotent Stem Cells of Both Fallopian Tube and Skin Origins.

机构信息

Department of Pediatrics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Tzu Chi University, Hualien 97004, Taiwan.

Department of Orthopedics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Tzu Chi University, Hualien 97004, Taiwan.

出版信息

Cells. 2023 Nov 16;12(22):2635. doi: 10.3390/cells12222635.

Abstract

Fallopian tube epithelial cells (FTECs) play a significant role in the development of high-grade serous ovarian cancer (HGSOC), but their utilization in in vitro experiments presents challenges. To address these limitations, induced pluripotent stem cells (iPSCs) have been employed as a potential solution, driven by the hypothesis that orthologous iPSCs may offer superior differentiation capabilities compared with their non-orthologous counterparts. Our objective was to generate iPSCs from FTECs, referred to as FTEC-iPSCs, and compare their differentiation potential with iPSCs derived from skin keratinocytes (NHEK). By introducing a four-factor Sendai virus transduction system, we successfully derived iPSCs from FTECs. To assess the differentiation capacity of iPSCs, we utilized embryoid body formation, revealing positive immunohistochemical staining for markers representing the three germ layers. In vivo tumorigenesis evaluation further validated the pluripotency of iPSCs, as evidenced by the formation of tumors in immunodeficient mice, with histological analysis confirming the presence of tissues from all three germ layers. Quantitative polymerase chain reaction (qPCR) analysis illuminated a sequential shift in gene expression, encompassing pluripotent, mesodermal, and intermediate mesoderm-related genes, during the iPSC differentiation process into FTECs. Notably, the introduction of WNT3A following intermediate mesoderm differentiation steered the cells toward a FTEC phenotype, supported by the expression of FTEC-related markers and the formation of tubule-like structures. In specific culture conditions, the expression of FTEC-related genes was comparable in FTECs derived from FTEC-iPSCs compared with those derived from NHEK-iPSCs. To conclude, our study successfully generated iPSCs from FTECs, demonstrating their capacity for FTEC differentiation. Furthermore, iPSCs originating from orthologous cell sources exhibited comparable differentiation capabilities. These findings hold promise for using iPSCs in modeling and investigating diseases associated with these specific cell types.

摘要

输卵管上皮细胞(FTECs)在高级别浆液性卵巢癌(HGSOC)的发展中起着重要作用,但在体外实验中利用它们存在挑战。为了解决这些限制,诱导多能干细胞(iPSCs)已被用作一种潜在的解决方案,其假设是同源 iPSCs 可能提供比非同源对应物更好的分化能力。我们的目标是从输卵管上皮细胞(FTEC)中生成 iPSCs,称为 FTEC-iPSCs,并将其与从皮肤角质形成细胞(NHEK)中衍生的 iPSCs 的分化潜力进行比较。通过引入四因素 Sendai 病毒转导系统,我们成功地从 FTEC 中衍生出 iPSCs。为了评估 iPSCs 的分化能力,我们利用胚状体形成,阳性免疫组织化学染色标记代表三个胚层的标志物。体内肿瘤发生评估进一步验证了 iPSCs 的多能性,因为免疫缺陷小鼠形成肿瘤,组织学分析证实存在来自所有三个胚层的组织。定量聚合酶链反应(qPCR)分析揭示了在 iPSC 分化为 FTEC 的过程中基因表达的顺序变化,包括多能性、中胚层和中间中胚层相关基因。值得注意的是,在中间中胚层分化后引入 WNT3A 促使细胞向 FTEC 表型发展,这得到了 FTEC 相关标志物的表达和管状结构的形成的支持。在特定的培养条件下,FTEC-iPSC 衍生的 FTEC 相关基因的表达与 NHEK-iPSC 衍生的 FTEC 相关基因的表达相当。总之,我们的研究成功地从 FTEC 中生成了 iPSCs,证明了它们分化为 FTEC 的能力。此外,源自同源细胞来源的 iPSCs 表现出相当的分化能力。这些发现为使用 iPSCs 对与这些特定细胞类型相关的疾病进行建模和研究提供了希望。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c047/10670511/d42a5c622bd2/cells-12-02635-g001.jpg

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