Experimental murine models are an essential tool in the field of bone marrow (BM) transplantation research. Therefore, numerous mice are required to obtain a sufficient number of BM cells, which is in contrast with the Reduction principle of the 3R principles. The selection of the cell source and the isolation protocol are therefore critical in obtaining a sufficient yield of cells for experiments. Nowadays, the vertebrae are already used as an extra source of BM cells to enrich the number of isolated cells from the long bones and ilia (LBI), when needed. Yet, little is known if BM cells from LBI and vertebrae share the same characteristics and can be pooled together for further analysis. Therefore, in this study, we aimed to compare the quantity and characteristics of haematopoietic and stromal cell lines in the BM from the LBI and vertebrae. To count haematopoietic and mesenchymal stem/stromal progenitors, colony-forming unit assays were performed. To determine the expansion capacity of mesenchymal stem/stromal cells (MSCs), cultivation of MSCs and measurement of the expression of surface markers by flow cytometry was performed. The characterisation and enumeration of immune cell populations was also performed by flow cytometry. Here, we show that the vertebrae are a comparable source of BM cells to the LBI regarding the analysed parameters.

中文翻译：

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3R 原则的应用：椎骨作为鼠骨髓细胞的额外来源。

实验鼠模型是骨髓 (BM) 移植研究领域的重要工具。因此，需要大量小鼠才能获得足够数量的 BM 细胞，这与 3R 原理的还原原理相反。因此，细胞来源的选择和隔离协议对于获得足够的细胞产量进行实验至关重要。如今，椎骨已被用作 BM 细胞的额外来源，以在需要时丰富来自长骨和髂骨 (LBI) 的分离细胞的数量。然而，对于来自 LBI 和椎骨的 BM 细胞是否具有相同的特征并且可以汇集在一起​​以进行进一步分析，我们知之甚少。因此，在本研究中，我们旨在比较来自 LBI 和椎骨的 BM 中造血和基质细胞系的数量和特征。为了计数造血和间充质干/基质祖细胞，进行了集落形成单位测定。为了确定间充质干/基质细胞 (MSCs) 的扩增能力，进行了 MSCs 的培养和通过流式细胞术测量表面标志物的表达。免疫细胞群的表征和计数也通过流式细胞术进行。在这里，我们表明，就分析的参数而言，椎骨是与 LBI 相当的 BM 细胞来源。免疫细胞群的表征和计数也通过流式细胞术进行。在这里，我们表明，就分析的参数而言，椎骨是与 LBI 相当的 BM 细胞来源。免疫细胞群的表征和计数也通过流式细胞术进行。在这里，我们表明，就分析的参数而言，椎骨是与 LBI 相当的 BM 细胞来源。