Fennema Eelco M, Renard Auke J S, Leusink Anouk, van Blitterswijk Clemens A, de Boer Jan
Institute of Biomedical Technology, University of Twente, the Netherlands.
Acta Orthop. 2009 Oct;80(5):618-21. doi: 10.3109/17453670903278241.
Large inter-donor differences exist in human mesenchymal stem cell (hMSC) yield and the response of these cells to osteogenic stimuli. The source of these differences may be clinical differences in stem cell characteristics between individuals or the aspiration procedure itself.
From a total of 23 donors, we aimed to take 2 consecutive 10-mL aspirates from the same site in 17 donors and in 6 donors we aimed to take a 5-mL and a 20-mL aspirate. The aspiration was stopped either when the syringe was full or when no more bone marrow came through. Mononuclear cell yield (MNC), MSC yield, and differentiation capacity were analyzed for intra-donor and inter-donor variation. We analyzed the effect of the dilution with peripheral blood by drawing 20 mL at once.
There was a high correlation between the first and second aspiration volumes, and aspirates with a volume of less than 8 mL showed a large variation in cellular yield. The second 10-mL aspirate, and also 20-mL aspirates, contained a lower concentration of nucleated cells and yielded lower numbers of mesenchymal stem cells. No effect of the aspiration procedure on the biological characteristics of the mesenchymal stem cells was seen.
We recommend collection volumes of bone marrow aspirates of at least 8 mL to reduce the risk of obtaining aspirates with low cell numbers. From the same site, a second aspiration or an aspirate of > 10 mL can be drawn without any loss of biological quality due to dilution with peripheral blood.
人间充质干细胞（hMSC）的产量以及这些细胞对成骨刺激的反应在不同供体之间存在很大差异。这些差异的来源可能是个体间干细胞特征的临床差异或抽吸过程本身。
我们旨在从23名供体中，在17名供体的同一部位连续采集两份10毫升的抽吸物，在6名供体中分别采集一份5毫升和一份20毫升的抽吸物。当注射器装满或不再有骨髓流出时停止抽吸。分析单核细胞产量（MNC）、间充质干细胞产量和分化能力的供体内和供体间差异。我们通过一次性抽取20毫升血液来分析外周血稀释的影响。
第一次和第二次抽吸量之间存在高度相关性，体积小于8毫升的抽吸物细胞产量变化很大。第二次10毫升抽吸物以及20毫升抽吸物中，有核细胞浓度较低，间充质干细胞产量也较低。未观察到抽吸过程对间充质干细胞生物学特性有影响。
我们建议骨髓抽吸物的采集量至少为8毫升，以降低获得低细胞数抽吸物的风险。从同一部位可以进行第二次抽吸或抽取超过10毫升的抽吸物，而不会因外周血稀释而导致生物学质量有任何损失。
