Clonal expansion of adult rat hepatic stem cell lines by suppression of asymmetric cell kinetics (SACK)
Clonal expansion of adult rat hepatic stem cell lines by suppression of asymmetric cell kinetics (SACK)
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DOI：
10.1002/bit.10727
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发表时间：
2003-09-30
影响因子：
3.8
通讯作者：
Sherley, JL
中科院分区：
文献类型：
--
作者：
Lee, HS;Crane, GG;Sherley, JL
成体干细胞在生物医学领域具有潜在的应用价值，包括细胞替代疗法、基因疗法和组织工程。然而，由于在扩增功能性成体干细胞中遇到的困难，这种应用受到限制。我们开发了一种基于抑制不对称细胞动力学（SACK）的成人干细胞扩增问题的新方法。我们假设，不对称的细胞动力学，所需的成体干细胞功能，是一个主要的障碍，他们在文化中的扩展。因此，成体干细胞从不对称细胞动力学向对称细胞动力学的转化将促进其在培养物中的指数扩增和长期增殖。嘌呤核苷黄苷（Xs），促进鸟嘌呤核糖核苷酸的生物合成，可用于可逆地将细胞从不对称细胞动力学转化为对称细胞动力学。我们使用Xs补充剂从成年大鼠肝脏中获得具有成体肝干细胞特性的克隆上皮细胞系。两个Xs衍生的细胞系，Lig-8和Lig-13，的属性进行了详细描述，并与没有Xs补充的成年大鼠肝细胞系的属性进行了比较。Xs衍生的细胞系表现出Xs依赖性不对称细胞动力学和成熟肝分化标志物的Xs依赖性表达。有趣的是，Lig-8细胞产生具有与肝细胞分化一致的特性的后代，而Lig-13后代细胞具有与胆管上皮分化一致的特性。一个稳定的成人胆管细胞干细胞系以前没有描述过。与它们的衍生原理一致，SACK衍生的肝细胞系既不表现出衰老也不表现出致瘤性，并且它们的分化特性在长期培养后是稳定的。SACK衍生的干细胞系的这些特征强调了不对称的细胞动力学作为一种基本的成体干细胞特性，其具有成为扩增和增殖多种成体干细胞的一般方法的基础的潜力。(C)2003 Wiley Periodicals，Inc.
Adult stem cells have potential use for several biomedical applications, including cell replacement therapy, gene therapy, and tissue engineering. However, such applications have been limited due to difficulties encountered in expanding functional adult stem cells. We have developed a new approach to the problem of adult stem cell expansion based on the suppression of asymmetric cell kinetics (SACK). We postulated that asymmetric cell kinetics, required for adult stem cell function, were a major barrier to their expansion in culture. As such, conversion of adult stem cells from asymmetric cell kinetics to symmetric cell kinetics would promote their exponential expansion and longterm propagation in culture. The purine nucleoside xanthosine (Xs), which promotes guanine ribonucleotide biosynthesis, can be used to reversibly convert cells from asymmetric cell kinetics to symmetric cell kinetics. We used Xs supplementation to derive clonal epithelial cell lines from adult rat liver that have properties of adult hepatic stem cells. The properties of two Xs-derived cell lines, Lig-8 and Lig-13, are described in detail and compared to properties of adult rat hepatic cell lines derived without Xs supplementation. The Xs-derived cell lines exhibit Xs-dependent asymmetric cell kinetics and Xs-dependent expression of mature hepatic differentiation markers. Interestingly, Lig-8 cells produce progeny with properties consistent with hepatocyte differentiation, while Lig-13 progeny cells have properties consistent with bile duct epithelium differentiation. A stable adult cholangiocyte stem cell line has not been previously described. Consistent with the principles of their derivation, the SACK-derived hepatic cell lines exhibit neither senescence nor tumorigenic properties, and their differentiation properties are stable after longterm culture. These characteristics of SACK-derived stem cell lines underscore asymmetric cell kinetics as an essential adult stem cell property with potential to be the basis for a general approach to expansion and propagation of diverse adult stem cells. (C) 2003 Wiley Periodicals, Inc.