Laboratory for Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig Maximilians University, Nussbaumstr. 20, 80336, Munich, Germany.
Vet Res Commun. 2011 Dec;35(8):487-99. doi: 10.1007/s11259-011-9492-8. Epub 2011 Jul 15.
Adult stem cells are of particular interest for the therapeutic approach in the field of regenerative medicine. Due to their ease of harvest, adipose-derived mesenchymal stem cells (ASCs) are an attractive stem cell source that has become increasingly popular. Critical aspects of applied cell therapies are the circumstances of transport from the laboratory towards the site of operation and cell delivery into the desired area. With regard to these issues, agarose-hydrogel was analyzed as a cell carrier matrix of equine and canine ASCs in vitro, which can be used for minimally invasive application. Isolated ASCs were expanded and 2.5 × 10(6) cells were combined with agarose-hydrogel to build a 0.4% hydrogel-cell solution which was stored at two temperatures (room temperature (RT) vs. 37 °C). Cell viability was investigated (live-dead assay) at different time points (0, 1, 6 and 24 h) in order to determine i) the effect of different temperatures on the cell survival as well as ii) the maximum possible time span before implantation. CFU-assay and WST-1 assay were performed after 24 h incubation in agarose-hydrogel and the cells were induced into adipogenic and osteogenic differentiation to analyze the effects of the incubation on the cell behaviour. No negative effect of the agarose-hydrogel incubation was determined on the different species' cell behaviour at either RT or 37 °C with any of the assays used. We can recommend agarose-hydrogel as a cell carrier for cell implantation with a storage period of up to 24 h at room temperature or at 37 °C prior to implantation.
成体干细胞在再生医学领域的治疗方法中特别受到关注。由于其易于采集，脂肪来源的间充质干细胞（ASCs）是一种有吸引力的干细胞来源，越来越受到关注。应用细胞治疗的关键方面是从实验室到手术部位的运输情况和细胞递送到所需区域。关于这些问题，琼脂糖水凝胶被分析为马和犬 ASC 的细胞载体基质，可用于微创应用。分离的 ASC 被扩增，并且 2.5×10(6)个细胞与琼脂糖水凝胶结合以构建 0.4%的水凝胶-细胞溶液，将其储存在两个温度下（室温（RT）与 37°C）。在不同时间点（0、1、6 和 24 h）进行细胞活力研究（死活测定），以确定 i）不同温度对细胞存活的影响，以及 ii）植入前的最大可能时间范围。在琼脂糖水凝胶中孵育 24 h 后进行 CFU 测定和 WST-1 测定，并诱导细胞向成脂和成骨分化，以分析孵育对细胞行为的影响。使用任何一种测定方法，都没有确定琼脂糖水凝胶孵育对不同物种细胞行为在 RT 或 37°C 下的任何一种测定方法的负面影响。我们可以推荐琼脂糖水凝胶作为细胞植入的细胞载体，在室温下储存长达 24 小时，或在植入前在 37°C 下储存。
