Maintenance of differentiation potential of human bone marrow mesenchymal stem cells immortalized by human telomerase reverse transcriptase gene in despite of extensive proliferation
Maintenance of differentiation potential of human bone marrow mesenchymal stem cells immortalized by human telomerase reverse transcriptase gene in despite of extensive proliferation
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DOI：
10.1016/j.bbrc.2004.11.059
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发表时间：
2005-01-21
影响因子：
3.1
通讯作者：
Kassem, M
中科院分区：
文献类型：
--
作者：
Abdallah, BM;Haack-Sorensen, M;Kassem, M
人骨髓间充质干细胞（hMSC）是一种能够分化成多个谱系的干细胞。然而，在长期体外培养过程中，这些细胞表现出衰老相关的生长停滞和表型变化。我们最近证明，在hMSC中过度表达人类端粒酶逆转录酶（hTERT）可重建端粒酶活性并延长细胞寿命[Nat. biotechnology . 20(2002) 592]。在本研究中，我们基于不同的培养密度(1:2 (hMSC-TERT2), 1:4 （hMSC-TERT4）和1:20 (hMSC-TERT20)，对来自亲本hMSC-TERT细胞系的三个独立细胞系进行了广泛的表征。这3种细胞系在形态和生长速度上存在差异，但在3年的培养过程中，它们都保持了干细胞自我更新的特征，并能够分化为多种中胚层型细胞系：成骨细胞、脂肪细胞、软骨细胞和内皮样细胞。此外，使用流式细胞术进行的表面标记研究显示了与正常hMSC相似的模式。因此，通过hTERT过表达对hMSC进行端粒化可以维持hMSC的干细胞表型，这可能是获得足够数量的具有稳定表型的细胞的有用工具，可用于细胞分化机制研究和组织工程方案。(C) 2004爱思唯尔公司版权所有。
Human bone marrow mesenchymal stem cells (hMSC) represent a population of stem cells that are capable of differentiation into multiple lineages. However, these cells exhibit senescence-associated growth arrest and phenotypic changes during long-term in vitro culture. We have recently demonstrated that overexpression of human telomerase reverse transcriptase (hTERT) in hMSC reconstitutes telomerase activity and extends life span of the cells [Nat. Biotechnol. 20 (2002) 592]. In the present study, we have performed extensive characterization of three independent cell lines derived from the parental hMSC-TERT cell line based on different plating densities during expansion in culture: 1:2 (hMSC-TERT2), 1:4 (hMSC-TERT4), and 1:20 (hMSC-TERT20). The 3 cell lines exhibited differences in morphology and growth rates but they all maintained the characteristics of self-renewing stem cells and the ability to differentiate into multiple mesoderm-type cell lineages: osteoblasts, adipocytes, chondrocytes, and endothelial-like cells over a 3-year period in culture. Also, surface marker studies using flow cytometry showed a pattern similar to that known from normal hMSC. Thus, telomerization of hMSC by hTERT overexpression maintains the stem cell phenotype of hMSC and it may be a useful tool for obtaining enough number of cells with a stable phenotype for mechanistic studies of cell differentiation and for tissue engineering protocols. (C) 2004 Elsevier Inc. All rights reserved.