Oxygen tension modifies the 'stemness' of human cord blood-derived stem cells
Oxygen tension modifies the 'stemness' of human cord blood-derived stem cells
复制标题
DOI：
10.3109/14653249.2012.671518
复制
发表时间：
2012-09-01
期刊：
影响因子：
4.5
通讯作者：
Koegler, Gesine
中科院分区：
文献类型：
--
作者：
Buchheiser, Anja;Houben, Amelie Pia;Koegler, Gesine
背景目标。在源自人脐带血（CB）的不同干细胞群中，非限制性成体干细胞（USSC）通过同源框（HOX）基因的表达模式、delta-like1同源物（DLK1）表达和成脂分化潜力与CB间充质基质细胞（CB MSC）区分开来。在这项研究中，我们研究了氧张力对干细胞标记基因的生成、增殖和表达的影响，这在临床应用的大规模细胞培养过程中可能至关重要。方法。我们在 5% 和 20% O-2 下培养 CB 衍生干细胞。我们分析了端粒长度的缩短，并使用逆转录 (RT)-聚合酶链式反应 (PCR) 和实时 PCR 来研究基因表达。此外，我们还进行了成脂和成骨的体外分化。结果。将 USSC 或 CB MSC 的培养条件从 20% O-2 改为 5% O-2 没有显着影响。相比之下，源自在 5% O-2 下制备的原代培养物的细胞群既不符合 USSC 也不符合 CB MSC。当转化为 20% 时，它们的增殖减弱，端粒缩短加速，六种细胞系中的两种停止表达 HOX 基因。其他细胞群的 HOX 代码不受培养条件的影响。结论。在生成过程中改变培养条件可能会影响细胞特性，例如 HOX 代码。在处理临床应用的细胞培养物时需要考虑这些影响。
Background aims. Amongst different stem cell populations derived from human cord blood (CB), unrestricted somatic stem cells (USSC) are distinguished from CB mesenchymal stromal cells (CB MSC) by expression patterns of homeobox (HOX) genes, delta-like1 homolog (DLK1) expression and adipogenic differentiation potential. In this study we investigated the effects of oxygen tension on the generation, proliferation and expression of stem cell marker genes, which could be critical during large-scale cell culture for clinical applications. Methods. We cultured CB-derived stem cells at 5% and 20% O-2. Telomere length shortening was analyzed and we investigated gene expression using reverse-transcription (RT)-polymerase chain reaction (PCR) and real-time PCR. Additionally we performed adipogenic and osteogenic in vitro differentiation. Results. Altering the cultivation conditions of USSC or CB MSC from 20% to 5% O-2 had no significant impact. In contrast, cell populations derived from primary cultures prepared at 5% O-2 qualified as neither USSC nor as CB MSC. When converted to 20%, their proliferation was diminished, telomere shortening was accelerated, and two of six cell lines ceased expression of HOX genes. The HOX code of the other cell populations was not been affected by culture conditions. Conclusions. Altering culture conditions during generation can impact cell characteristics such as the HOX code. These effects need to be considered when dealing with cell cultures for clinical applications.