Functional modulation of ES-derived hepatocyte lineage cells via substrate compliance alteration
Functional modulation of ES-derived hepatocyte lineage cells via substrate compliance alteration
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DOI：
10.1007/s10439-008-9458-3
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发表时间：
2008-05-01
影响因子：
3.8
通讯作者：
Langrana, Noshir A.
中科院分区：
文献类型：
--
作者：
Li, Lulu;Sharma, Nripen;Langrana, Noshir A.
多能胚胎干细胞代表了一种有前途的可再生细胞来源，可产生多种分化的细胞类型，包括肝细胞谱系细胞，并可能最终被纳入体外生物人工肝装置和细胞替代疗法。最近，我们和其他人利用丁酸钠直接分化肝细胞样细胞从小鼠胚胎干细胞培养在单层配置。然而，为了将干细胞技术纳入临床和药学应用，并希望增加这些分化细胞用于肝病治疗的治疗潜力，在其二次培养环境中维持分化功能持续较长时间仍然是一个主要挑战。在目前的工作中，我们已经研究了使用聚丙烯酰胺水凝胶与定义的机械顺应性作为细胞培养平台，用于改善和/或稳定这些肝细胞样细胞的功能。几种功能测定，例如，尿素分泌、细胞内白蛋白含量和白蛋白分泌，以表征在刚度为5、46.6和230 kPa的聚丙烯酰胺凝胶上的细胞的肝功能。结合机械和细胞形态学特征，我们发现丁酸钠分化细胞的肝功能在顺应性基质上持续并进一步增强。这项研究有望提供通过机械刺激调节干细胞分化的见解，并帮助我们设计各种动态培养系统，用于组织和细胞工程。
Pluripotent embryonic stem cells represent a promising renewable cell source to generate a variety of differentiated cell types including hepatocyte lineage cells, and may ultimately be incorporated into extracorporeal bioartificial liver devices and cell replacement therapies. Recently, we and others have utilized sodium butyrate to directly differentiate hepatocyte-like cells from murine embryonic stem cells cultured in a monolayer configuration. However, to incorporate stem cell technology into clinical and pharmaceutical applications, and hopefully increase the therapeutic potential of these differentiated cells for liver disease treatment, a major challenge remains in sustaining differentiated functions for an extended period of time in their secondary culture environment. In the present work, we have investigated the use of polyacrylamide hydrogels with defined mechanical compliances as a cell culture platform for improving and/or stabilizing functions of these hepatocyte-like cells. Several functional assays, e.g., urea secretion, intracellular albumin content, and albumin secretion, were performed to characterize hepatic functions of cells on polyacrylamide gels with stiffnesses of 5, 46.6, and 230 kPa. In conjunction with the mechanical and cell morphological characterization, we showed that hepatic functions of sodium butyrate differentiated cells were sustained and further enhanced on compliant substrates. This study promises to offer insights into regulating stem cell differentiation via mechanical stimuli, and assist us with designing a variety of dynamic culture systems for applications in tissue and cellular engineering.