利用高通量表达谱和下一代知识发现平台表征人类脐带血来源的肥大细胞

IF 2.8 4区 医学 Q2 PATHOLOGY Experimental and molecular pathology Pub Date : 2023-08-01 DOI:10.1016/j.yexmp.2023.104867
Sherin Bakhashab , Ghalya H. Banafea , Farid Ahmed , Haneen Alsehli , Huda F. AlShaibi , Nadia Bagatian , Ohoud Subhi , Kalamegam Gauthaman , Mahmood Rasool , Hans-Juergen Schulten , Peter Natesan Pushparaj
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引用次数: 0

摘要

肥大细胞(MC)是组织固有的先天免疫细胞,表达免疫球蛋白E的高亲和力受体,负责宿主防御和一系列与免疫系统相关的疾病。在本研究中,我们旨在使用下一代知识发现方法,与来源于CD34祖细胞的细胞相比,表征人脐血来源的MCs(hCBMCs)的途径和基因特征。使用磁性活化细胞分选从人脐带血中分离CD34+细胞,并在补充rhIL-6和rhSCF的情况下分化为MCs 6-8周。通过流式细胞术分析hCBMCs的纯度,显示表面标记CD117+CD34−CD45−CD23−FcεR1αdim。从hCBMCs和CD34−细胞中分离总RNA,并使用微阵列进行杂交。使用iPathway指南和预排序基因集富集分析来分析差异表达的基因。下一代知识发现平台揭示了hCBMCs中富集的MC特异性基因特征和分子途径,并涉及免疫反应库。
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Characterization of human umbilical cord blood-derived mast cells using high-throughput expression profiling and next-generation knowledge discovery platforms

Mast cells (MCs) are tissue-resident innate immune cells that express the high-affinity receptor for immunoglobulin E and are responsible for host defense and an array of diseases related to immune system. We aimed in this study to characterize the pathways and gene signatures of human cord blood-derived MCs (hCBMCs) in comparison to cells originating from CD34 progenitors using next-generation knowledge discovery methods. CD34+ cells were isolated from human umbilical cord blood using magnetic activated cell sorting and differentiated into MCs with rhIL-6 and rhSCF supplementation for 6–8 weeks. The purity of hCBMCs was analyzed by flow cytometry exhibiting the surface markers CD117+CD34CD45CD23FcεR1αdim. Total RNA from hCBMCs and CD34 cells were isolated and hybridized using microarray. Differentially expressed genes were analyzed using iPathway Guide and Pre-Ranked Gene Set Enrichment Analysis. Next-generation knowledge discovery platforms revealed MC-specific gene signatures and molecular pathways enriched in hCBMCs and pertain the immunological response repertoire.

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来源期刊
CiteScore
8.90
自引率
0.00%
发文量
78
审稿时长
11.5 weeks
期刊介绍: Under new editorial leadership, Experimental and Molecular Pathology presents original articles on disease processes in relation to structural and biochemical alterations in mammalian tissues and fluids and on the application of newer techniques of molecular biology to problems of pathology in humans and other animals. The journal also publishes selected interpretive synthesis reviews by bench level investigators working at the "cutting edge" of contemporary research in pathology. In addition, special thematic issues present original research reports that unravel some of Nature''s most jealously guarded secrets on the pathologic basis of disease. Research Areas include: Stem cells; Neoangiogenesis; Molecular diagnostics; Polymerase chain reaction; In situ hybridization; DNA sequencing; Cell receptors; Carcinogenesis; Pathobiology of neoplasia; Complex infectious diseases; Transplantation; Cytokines; Flow cytomeric analysis; Inflammation; Cellular injury; Immunology and hypersensitivity; Athersclerosis.
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