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[层粘连蛋白基因家族在猪多能干细胞中的表达]

[Expression of Laminin gene family in porcine pluripotent stem cells].

作者信息

Li Huan, Yu Tong, Ma Yangyang, Wang Huayan

机构信息

Shaanxi Center for Stem Cell Engineering and Technology, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2017 Aug 25;33(8):1304-1314. doi: 10.13345/j.cjb.170101.

Abstract

Laminin (LN) proteins are important components of extracellular matrix. These proteins regulate cell proliferation, differentiation, migration, and tissue repair. The LN family has 12 genes that encode 5 α, 4 β, and 3 γ proteins. LamininA5 (LAMA5) as an important gene can support pluripotent cell growth and have been widely studied. However, porcine LAMA5 is absent in all tested porcine genomic databases so far. In this study, we confirmed for the first time the existence of porcine LAMA5 through bioinformatics analysis, and verified this result by cDNA cloning and sequencing. To reveal the expression pattern of Laminin gene family, we detected the expression of Laminin genes in porcine tissues, somatic cells, and porcine induced pluripotent stem cells (piPSCs). The results showed that an alternative splicing variant of Laminin B1 (LAMB1-a) was found exclusively in all tested piPSCs. The expression of this alternative splicing variant is positively correlated with the pluripotent state of piPSCs. The above findings provide evidences and foundations for the father use of LN as extracellular matrix to facilitate the derivation and culture of porcine pluripotent stem cells.

摘要

层粘连蛋白(LN)是细胞外基质的重要组成部分。这些蛋白质调节细胞增殖、分化、迁移和组织修复。层粘连蛋白家族有12个基因,编码5种α、4种β和3种γ蛋白质。层粘连蛋白A5(LAMA5)作为一个重要基因,能够支持多能细胞生长,并且已经得到广泛研究。然而,到目前为止,在所有测试的猪基因组数据库中均未发现猪LAMA5。在本研究中,我们首次通过生物信息学分析证实了猪LAMA5的存在,并通过cDNA克隆和测序验证了这一结果。为了揭示层粘连蛋白基因家族的表达模式,我们检测了层粘连蛋白基因在猪组织、体细胞和猪诱导多能干细胞(piPSC)中的表达。结果表明,在所有测试的piPSC中仅发现了层粘连蛋白B1的一种可变剪接变体(LAMB1-a)。这种可变剪接变体的表达与piPSC的多能状态呈正相关。上述发现为进一步利用LN作为细胞外基质促进猪多能干细胞的诱导和培养提供了证据和基础。

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