METHODS: In this study, human pluripotent stem cell-derived HSCs (scHSCs), benchmarked to human primary HSC, were exposed to 48-hour starvation of retinol (ROL) and palmitic acid (PA) in the presence or absence of the potent HSC activator TGF-β. The interventions were studied by an extensive set of phenotypic and functional analyses, including transcriptomic analysis, measurement of activation-related proteins and cytokines, VA- and LD storage, and cell energy metabolism.
RESULTS: The results show that though the starvation of ROL and PA alone did not induce scHSC activation, the starvation amplified the TGF-β-induced activation-related transcriptome. However, TGF-β-induced activation alone did not lead to a reduction in VA or LD stores. Additionally, reduced glycolysis and increased mitochondrial fission were observed in response to TGF-β.
CONCLUSIONS: scHSCs are robust models for activation studies. The loss of VA and LDs is not sufficient for scHSC activation in vitro, but may amplify the TGF-β-induced activation response. Collectively, our work provides an extensive framework for studying human HSCs in healthy and diseased conditions.
方法:在本研究中,人多能干细胞来源的HSC(scHSC),以人类初级HSC为基准,在存在或不存在有效的HSC激活剂TGF-β的情况下,暴露于视黄醇(ROL)和棕榈酸(PA)的48小时饥饿。通过广泛的表型和功能分析研究了干预措施,包括转录组学分析,激活相关蛋白和细胞因子的测量,VA和LD存储,和细胞能量代谢。
结果:结果表明,尽管单独的ROL和PA饥饿并不诱导scHSC活化,饥饿放大了TGF-β诱导的活化相关转录组。然而,单独TGF-β诱导的激活不会导致VA或LD存储的减少。此外,对TGF-β的反应观察到糖酵解减少和线粒体裂变增加。
结论:scHSC是激活研究的稳健模型。VA和LD的损失不足以在体外激活scHSC,但可能会放大TGF-β诱导的激活反应。总的来说,我们的工作为在健康和疾病条件下研究人类HSC提供了一个广泛的框架.