Autologous Serum Improves Yield and Metabolic Capacity of Monocyte-Derived Hepatocyte-Like Cells: Possible Implication for Cell Transplantation
Autologous Serum Improves Yield and Metabolic Capacity of Monocyte-Derived Hepatocyte-Like Cells: Possible Implication for Cell Transplantation
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DOI：
10.3727/096368910x550224
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发表时间：
2011-01-01
影响因子：
3.3
通讯作者：
Nuessler, A. K.
中科院分区：
文献类型：
--
作者：
Ehnert, S.;Seeliger, C.;Nuessler, A. K.
肝细胞移植是一种治疗各种急慢性肝病的方法。由于原代细胞的可用性有限，因此对肝细胞样细胞（例如，由外周血单核细胞产生的新肝细胞）。本研究的目的是评估六种不同的人AB血清、胎牛血清或自体血清对新生肝细胞产生的影响。新生肝细胞的产量和质量根据不同的血清而变化很大。在整个生产过程中使用自体血清，我们不断产生最高量的具有I期最高代谢活性的细胞（例如，CYP 1A 1/2、CYP 3A 4）和II相酶（例如，谷胱甘肽-S-转移酶）。此外，在检查葡萄糖和尿素代谢时也观察到类似的效果。特别是葡萄糖-6-磷酸酶和PAS染色显示出明显的血清依赖性差异。通过测量单核细胞来源的可编程细胞（PCMO）中TNF-α、TGF-β和RANKL的分泌、MMP活性以及不同白介素的mRNA水平来研究巨噬细胞活化的作用。我们的数据清楚地表明，使用自体血清减少了PCMO中的初始巨噬细胞活化，随后提高了分化的新肝细胞的产量和功能。这里提出的自体方法也可能在其他干细胞制备过程中有用，其中在生成期间细胞活化应保持在最低限度。
Hepatocyte-transplantation is a therapeutic approach for diverse acute and chronic liver diseases. As availability of primary cells is limited, there is an increasing demand for hepatocyte-like cells (e.g., neohepatocytes generated from peripheral blood monocytes). The aim of this study was to evaluate the effects of six different human AB sera, fetal calf serum, or autologous serum on production of neohepatocytes. The yield and quality of neohepatocytes varied considerably depending on the different sera. Using autologous sera for the whole production process we constantly generated the highest amount of cells with the highest metabolic activity for phase I (e.g., CYP1A1/2, CYP3A4) and phase II enzymes (e.g., glutathione-S-transferase). Moreover, similar effects were seen examining glucose and urea metabolism. Especially, glucose-6-phosphatase and PAS staining showed distinct serum-dependent differences. The role of macrophage activation was investigated by measuring the secretion of TNF-alpha, TGF-beta, and RANKL, MMP activity, as well as mRNA levels of different interleukins in programmable cells of monocytic origin (PCMO). Our data clearly demonstrate that the use of autologous serum reduced initial macrophage activation in PCMOs and subsequently improved both yield and function of differentiated neohepatocytes. The autologous approach presented here might also be useful in other stem cell preparation processes where cell activation during generation shall be kept to a minimum.