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绵羊脐带间充质干细胞的分离与鉴定

Isolation and characterization of ovine umbilical cord-derived mesenchymal stem cells.

作者信息

Zhao Sirguleng, Tao Li, Tian Yunyun, Tai Dapeng, Liu Pengxia, Liu Dongjun

机构信息

State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, Inner Mongolia, P.R. China.

出版信息

Cytotechnology. 2019 Feb;71(1):277-286. doi: 10.1007/s10616-018-0284-7. Epub 2019 Jan 2.

Abstract

Mesenchymal stem cells (MSCs) are able to self-renew and have multi-lineage differentiation potential. However, studies on ovine umbilical cord-derived MSCs (UC-MSCs) are limited. Our study aimed to isolate and characterize ovine UC-MSCs. We successfully isolated ovine UC-MSCs and defined their surface marker profile using immunofluorescence analysis. Ovine UC-MSCs were found to be positive for cell surface markers CD13, CD29, CD44, CD90, and CD106, and negative for cell surface marker CD45. Assessment of the proliferation potential of ovine UC-MSCs showed that from day 3 of cultivation a plateau phase was reached. And compare to passage 10, 15, 20 cells, passage 5 cells proliferating the fastest. Differentiation of ovine UC-MSCs into adipocytes, osteocytes, and chondrocytes was also demonstrated by staining for tissue-specific markers and using quantitative real-time polymerase chain reaction for specific marker gene expression. This study demonstrates the existence of a MSC population within the ovine umbilical cord, which maintained a normal karyotype up to passage 20.

摘要

间充质干细胞(MSCs)能够自我更新并具有多向分化潜能。然而,关于绵羊脐带间充质干细胞(UC-MSCs)的研究有限。我们的研究旨在分离并鉴定绵羊UC-MSCs。我们成功分离出绵羊UC-MSCs,并通过免疫荧光分析确定了它们的表面标志物谱。发现绵羊UC-MSCs的细胞表面标志物CD13、CD29、CD44、CD90和CD106呈阳性,而细胞表面标志物CD45呈阴性。对绵羊UC-MSCs增殖潜能的评估表明,从培养第3天起进入平台期。与第10、15、20代细胞相比,第5代细胞增殖最快。通过组织特异性标志物染色以及使用定量实时聚合酶链反应检测特定标志物基因表达,也证实了绵羊UC-MSCs可分化为脂肪细胞、骨细胞和软骨细胞。本研究证明了绵羊脐带中存在间充质干细胞群体,其核型在传至第20代时仍保持正常。

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