Servicio de Hematología, Hospital Universitario de Salamanca, Salamanca, Spain.
PLoS One. 2011;6(10):e26241. doi: 10.1371/journal.pone.0026241. Epub 2011 Oct 20.
The aim of the present study was to determine how mesenchymal stem cells (MSC) could improve bone marrow (BM) stroma function after damage, both in vitro and in vivo. Human MSC from 20 healthy donors were isolated and expanded. Mobilized selected CD34(+) progenitor cells were obtained from 20 HSCT donors. For in vitro study, long-term bone marrow cultures (LTBMC) were performed using a etoposide damaged stromal model to test MSC effect in stromal confluence, capability of MSC to lodge in stromal layer as well as some molecules (SDF1, osteopontin,) involved in hematopoietic niche maintenance were analyzed. For the in vivo model, 64 NOD/SCID recipients were transplanted with CD34+ cells administered either by intravenous (i.v.) or intrabone (i.b.) route, with or without BM derived MSC. MSC lodgement within the BM niche was assessed by FISH analysis and the expression of SDF1 and osteopontin by immunohistochemistry. In vivo study showed that when the stromal damage was severe, TP-MSC could lodge in the etoposide-treated BM stroma, as shown by FISH analysis. Osteopontin and SDF1 were differently expressed in damaged stroma and their expression restored after TP-MSC addition. Human in vivo MSC lodgement was observed within BM niche by FISH, but MSC only were detected and not in the contralateral femurs. Human MSC were located around blood vessels in the subendoestal region of femurs and expressed SDF1 and osteopontin. In summary, our data show that MSC can restore BM stromal function and also engraft when a higher stromal damage was done. Interestingly, MSC were detected locally where they were administered but not in the contralateral femur.
本研究旨在确定间充质干细胞（MSC）如何在体外和体内改善受损骨髓（BM）基质的功能。从 20 名健康供体中分离和扩增人 MSC。从 20 名 HSCT 供体中获得动员的选择 CD34（+）祖细胞。为了进行体外研究，使用依托泊苷损伤的基质模型进行长期骨髓培养（LTBMC），以测试 MSC 对基质汇合的影响、MSC 驻留在基质层中的能力以及一些参与造血龛维持的分子（SDF1、骨桥蛋白）。对于体内模型，将 64 名 NOD/SCID 受体用静脉内（i.v.）或骨内（i.b.）途径给予 CD34+细胞进行移植，并用或不用 BM 来源的 MSC。通过 FISH 分析评估 MSC 在 BM 龛内的定位，通过免疫组织化学分析 SDF1 和骨桥蛋白的表达。体内研究表明，当基质损伤严重时，TP-MSC 可以驻留在依托泊苷处理的 BM 基质中，如 FISH 分析所示。骨桥蛋白和 SDF1 在受损的基质中表达不同，添加 TP-MSC 后其表达得到恢复。通过 FISH 在体内观察到人类 MSC 驻留在 BM 龛内，但仅检测到 MSC，而不是在对侧股骨中。人类 MSC 位于股骨亚内皮区域的血管周围，表达 SDF1 和骨桥蛋白。总之，我们的数据表明，MSC 可以恢复 BM 基质的功能，并且在基质损伤更大时也可以植入。有趣的是，MSC 仅在给药部位局部检测到，而不在对侧股骨中检测到。
