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干细胞蛋白质组:源自脐带血的人间充质干细胞概况

Stem cell proteomes: a profile of human mesenchymal stem cells derived from umbilical cord blood.

作者信息

Feldmann Robert E, Bieback Karen, Maurer Martin H, Kalenka Armin, Bürgers Heinrich F, Gross Benjamin, Hunzinger Christian, Klüter Harald, Kuschinsky Wolfgang, Eichler Hermann

机构信息

Department of Physiology and Pathophysiology, University of Heidelberg, Heidelberg, Germany.

出版信息

Electrophoresis. 2005 Jul;26(14):2749-58. doi: 10.1002/elps.200410406.

Abstract

Multipotent mesenchymal stem cells (MSCs) derived from human umbilical cord blood (UCB) represent promising candidates for the development of future strategies in cellular therapy. To create a comprehensive protein expression profile for UCB-MSCs, one UCB unit from a full-term delivery was isolated from the unborn placenta, transferred into culture, and their whole-cell protein fraction was subjected to two-dimensional electrophoresis (2-DE). Unambiguous protein identification was achieved with peptide mass fingerprinting matrix-assisted laser desorption/ionization - time of flight - mass spectrometry (MALDI-TOF-MS), peptide sequencing (MALDI LIFT-TOF/TOF MS), as well as gel-matching with previously identified databases. In overall five replicate 2-DE runs, a total of 2037 +/- 437 protein spots were detected of which 205 were identified representing 145 different proteins and 60 isoforms or post-translational modifications. The identified proteins could be grouped into several functional categories, such as metabolism, folding, cytoskeleton, transcription, signal transduction, protein degradation, detoxification, vesicle/protein transport, cell cycle regulation, apoptosis, and calcium homeostasis. The acquired proteome map of nondifferentiated UCB-MSCs is a useful inventory which facilitates the identification of the normal proteomic pattern as well as its changes due to activated or suppressed pathways of cytosolic signal transduction which occur during proliferation, differentiation, or other experimental conditions.

摘要

源自人脐带血(UCB)的多能间充质干细胞(MSC)是细胞治疗未来策略开发中很有前景的候选者。为了创建UCB-MSC的全面蛋白质表达谱,从足月分娩的未出生胎盘中分离出一个UCB单元,转移到培养基中,并对其全细胞蛋白质组分进行二维电泳(2-DE)。通过肽质量指纹图谱基质辅助激光解吸/电离-飞行时间-质谱(MALDI-TOF-MS)、肽测序(MALDI LIFT-TOF/TOF MS)以及与先前鉴定的数据库进行凝胶匹配,实现了明确的蛋白质鉴定。在总共五次重复的2-DE运行中,共检测到2037±437个蛋白点,其中205个被鉴定,代表145种不同蛋白质以及60种异构体或翻译后修饰。鉴定出的蛋白质可分为几个功能类别,如代谢、折叠、细胞骨架、转录、信号转导、蛋白质降解、解毒、囊泡/蛋白质运输、细胞周期调控、细胞凋亡和钙稳态。所获得的未分化UCB-MSC蛋白质组图谱是一份有用的清单,有助于识别正常蛋白质组模式及其因细胞溶质信号转导途径在增殖、分化或其他实验条件下激活或抑制而发生的变化。

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