Discovery Biology, St. Louis Laboratories, Pfizer Global Research & Development, Pfizer Inc., Chesterfield, MO, USA.
Dig Dis Sci. 2012 May;57(5):1152-62. doi: 10.1007/s10620-012-2084-9. Epub 2012 Feb 23.
Hepatic stellate cells (HSC) play a major role in the progression of liver fibrosis.
The aim of our study was to investigate whether rat HSC cultured on a nanofiber membrane (NM) retain their quiescent phenotype during both short- and long-term culture and whether activated HSC revert to a quiescent form when re-cultured on NM.
Rat HSC cultured for 1 day on plastic plates (PP) were used as quiescent HSC, while cells cultured for 1 week on PP were considered to be activated HSC. Quiescent or activated HSC were subsequently plated on PP or NM and cultured for an additional 4 days at which time their gene expression, stress fiber development, and growth factor production were determined. For long-term culture, HSC were grown on NM for 20 days and the cells then replated on PP and cultured for another 10 days.
Expression of marker genes for HSC activation, stress fiber development, and growth factor production were significantly lower in both quiescent and activated HSC cultured on NM than in those cultured on PP. After long-term culture on NM, activation marker gene expression and stress fiber development were still significantly lower in HSC than in PP, and HSC still retained the ability to activate when replated onto PP.
HSC cultured on NM retained quiescent characteristics after both short- and long-term culture while activated HSC reverted toward a quiescent state when cultured on NM. Cultures of HSC grown on NM are a useful in vitro model to investigate the mechanisms of activation and deactivation.
肝星状细胞(HSC)在肝纤维化的进展中起着重要作用。
本研究旨在探讨在短期和长期培养过程中,培养在纳米纤维膜(NM)上的大鼠 HSC 是否能保持其静止表型,以及激活的 HSC 在重新培养在 NM 上时是否能恢复到静止状态。
培养在塑料板(PP)上 1 天的大鼠 HSC 用作静止 HSC,而培养在 PP 上 1 周的细胞被认为是激活的 HSC。随后将静止或激活的 HSC 分别接种在 PP 或 NM 上,并在培养 4 天后测定其基因表达、应激纤维的发育和生长因子的产生。进行长期培养时,将 HSC 在 NM 上培养 20 天,然后将细胞再接种在 PP 上,并培养 10 天。
与在 PP 上培养的静止和激活的 HSC 相比,在 NM 上培养的 HSC 的激活标志物基因表达、应激纤维发育和生长因子产生均显著降低。在 NM 上长期培养后,HSC 的激活标志物基因表达和应激纤维发育仍明显低于 PP,且当再接种在 PP 上时,HSC 仍具有激活的能力。
在短期和长期培养后,培养在 NM 上的 HSC 保留了静止特征,而激活的 HSC 在培养在 NM 上时则恢复到静止状态。在 NM 上生长的 HSC 培养物是研究激活和去激活机制的有用体外模型。