The Second Department of Craniomaxillofacial Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 33 Ba-Da-Chu Road, Shi-Jing-Shan District, Beijing, 100144, China.
Cleft Lip and Palate Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 33 Ba-Da-Chu Road, Shi-Jing-Shan District, Beijing, 100144, China.
Aesthetic Plast Surg. 2024 May;48(9):1855-1866. doi: 10.1007/s00266-024-03892-8. Epub 2024 Feb 22.
Bone marrow mononuclear cells (BMMNCs) have great potential in bone regenerative therapy. The main method used today to obtain BMMNCs is Ficoll density gradient centrifugation. However, the centrifugal force for this isolation method is still suboptimal.
To determine the optimal centrifugal force in Ficoll density gradient centrifugation of bone marrow (BM) to achieve high stem/progenitor cell content BMMNCs for regenerative therapy.
BM was aspirated from nine minipigs and divided into three groups according to different centrifugal forces (200 g, 300 g and 400 g). Immediately after BMMNCs were obtained from each group by Ficoll density gradient centrifugation, residual red blood cell (RBC) level, nucleated cell counting, viability and flow cytometric analyses of apoptosis and reactive oxygen species (ROS) generation were measured. The phenotypic CD90 and colony formation analyses of BMMNCs of each group were performed as well. Bone marrow-derived mesenchymal stem cells (BMSCs) were harvested at passage 2, then morphology, cell phenotype, proliferation, adipogenic, chondrogenic and osteogenic lineage differentiation potential of BMSCs from each group were compared.
The 300 g centrifugal force was able to isolate BMMNCs from BM with the same efficiency as 400 g and provided significantly higher yields of CD90 BMSCs and fibroblastic colony-forming units of BMSC (CFU-f(BMSC)), which is more crucial for the regenerative efficacy of BMMNCs. Meanwhile, 200 g hosted the most RBC contamination and minimum CFU-f (BMSC) yield, which will be disadvantageous for BMMNC-based cell therapy. As for in vitro cultured BMSCs which were isolated from BMMNCs by different centrifugal forces, no significant differences were found on morphology, cell proliferation rate, phenotypic marker, adipogenic, chondrogenic and osteogenic differentiation potential.
300 g may be the optimal centrifugal force when using Ficoll density gradient centrifugation to isolate BMMNCs for bone regenerative therapy.
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骨髓单核细胞（BMMNCs）在骨再生治疗中有很大的潜力。目前获得 BMMNCs 的主要方法是菲可密度梯度离心。然而，这种分离方法的离心力仍然不理想。
确定菲可密度梯度离心骨髓（BM）时的最佳离心力，以获得高含量的干细胞/祖细胞的 BMMNCs，用于再生治疗。
从 9 头小型猪中抽吸 BM，并根据不同的离心力（200g、300g 和 400g）分为三组。立即通过菲可密度梯度离心从每组中获得 BMMNCs 后，测量残留红细胞（RBC）水平、核细胞计数、活力和凋亡和活性氧（ROS）生成的流式细胞术分析。还对每组 BMMNCs 的表型 CD90 和集落形成分析进行了研究。收获第 2 代骨髓间充质干细胞（BMSCs），然后比较每组 BMSCs 的形态、细胞表型、增殖、成脂、成软骨和成骨谱系分化潜能。
300g 的离心力能够以与 400g 相同的效率从 BM 中分离 BMMNCs，并提供显著更高产量的 CD90 BMSCs 和纤维母细胞集落形成单位的 BMSC（CFU-f（BMSC）），这对 BMMNCs 的再生疗效更为重要。同时，200g 含有最多的 RBC 污染和最小的 CFU-f（BMSC）产量，这对基于 BMMNC 的细胞治疗不利。对于通过不同离心力从 BMMNCs 分离的体外培养的 BMSCs，在形态、细胞增殖率、表型标记物、成脂、成软骨和成骨分化潜能方面没有发现显著差异。
300g 可能是使用菲可密度梯度离心分离用于骨再生治疗的 BMMNCs 的最佳离心力。
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