Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University Medical Center, Düsseldorf, Germany.
Cytotherapy. 2012 Sep;14(8):967-82. doi: 10.3109/14653249.2012.671518. Epub 2012 Apr 12.
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.
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.
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.
在源自人脐带血（CB）的不同干细胞群体中，无限制体干细胞（USSC）通过同源盒（HOX）基因表达模式、delta-like1 同源物（DLK1）表达和脂肪生成分化潜力与 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志物基因生成、增殖和表达的影响，这在用于临床应用的大规模细胞培养过程中可能是至关重要的。
我们在 5%和 20%的 O(2)下培养 CB 衍生的干细胞。分析端粒缩短情况，并使用逆转录（RT）-聚合酶链反应（PCR）和实时 PCR 研究基因表达。此外，我们还进行了体外成脂和成骨分化。结果。将 USSC 或 CB MSC 的培养条件从 20%改变为 5% O(2)，没有显著影响。相比之下，从 5% O(2)制备的原代培养物衍生的细胞群体既不符合 USSC 也不符合 CB MSC。当转化为 20%时，它们的增殖减少，端粒缩短加速，并且六个细胞系中的两个停止表达 HOX 基因。其他细胞群体的 HOX 编码不受培养条件的影响。
在生成过程中改变培养条件会影响细胞特征，如 HOX 编码。在处理用于临床应用的细胞培养物时，需要考虑这些影响。
