Laminin111-based defined culture promoting self-renewing human pluripotent stem cells exhibiting properties of early post-implantation epiblast cells [Bulk_RNA_Seq]

In our study we sought to develop cultures highly enriched for self-renewing human pluripotent stem cells (hPSCs) with an early post-implantation phenotype, capturing the formative pluripotency phase in vitro. Our results demonstrate that hPSCs cultured on LN111 in defined conditions (LN-hPSCs) generate cultures highly enriched for genetically stable, self-renewing hPSCs exhibiting properties similar to the early-post implantation epiblast, including competence to give rise to the germ cell lineage. To analyze the global transcriptome of LN-hPSCs in comparison with conventional/primed and naive hPSCs, we performed RNA-seq of the three hPSC lines cultured under LN, primed, and naïve conditions. Overall design: Comparative gene expression profiling analysis of RNA-seq data for three independent hPSC lines HES-1 (ES01), HES-2 (ES02) (Reubinoff et al., 2000), and and H7 (WA07, NIH registry # 0061) (Thomson et al., 1998) cultured under LN, primed and naïve conditions.

本研究旨在构建高度富集自我更新型人类多能干细胞（human pluripotent stem cells，hPSCs）的培养体系，使其呈现早期植入后表型，从而在体外捕获成型多能性阶段。研究结果显示，在限定性培养体系中以层粘连蛋白111（LN111）为培养基质的hPSCs（LN-hPSCs）可形成高度富集遗传稳定、具备自我更新能力的hPSCs的培养物，其特性与早期植入后上胚层相似，包括具备分化为生殖细胞谱系的潜能。为对比分析LN-hPSCs与常规/始发态及初始态人类多能干细胞的全局转录组，我们对三种分别在LN111、始发态及初始态培养条件下培养的hPSC细胞系进行了RNA测序（RNA-seq）。整体实验设计：本研究对三种独立的hPSC细胞系——HES-1（ES01）、HES-2（ES02）（Reubinoff等，2000）以及H7（WA07，美国国立卫生研究院NIH注册编号#0061）（Thomson等，1998）——分别在LN111、始发态及初始态培养条件下进行培养，并对其RNA-seq数据开展比较基因表达谱分析。