层粘连蛋白基因家族在猪多能干细胞中的表达谱分析

Expression of Laminin gene family in porcine pluripotent stem cells

摘要:

层粘连蛋白 (Laminin) 是细胞外基质的重要成分，对细胞生长、分化、运动、组织修复和再生等发挥重要调节作用。Laminin包括有A1-5、B1-4和C1-3共12个编码基因，以不同的表达模式发挥功能。其中Laminin A5作为可以支持多能细胞生长的重要基因，得到广泛研究。但是，在所有猪相关数据库中，均未能查到Laminin A5的信息。文中通过生物信息学分析，首次确认了猪Laminin A5的存在，并进行了克隆和测序验证。为揭示Laminin基因家族在猪诱导多能干细胞 (iPSCs) 中的表达特性，检测了Laminin在猪各组织、体细胞和iPS细胞中的不同表达模式。发现Laminin B1基因在猪多能干细胞中存在特异性可变剪接体 (LAMB1-a)，且该可变剪切体的表达量与猪多能干细胞的重编程程度呈正相关。为进一步揭示和利用Laminin作为细胞外基质，用于猪多能干细胞的获取和培养优化奠定了基础。

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.