The potential use of bone marrow mesenchymal stromal cells (BM-MSCs) for the treatment of osteonecrosis in sickle cell disease (SCD) patients is increasing. However, convenient BM-MSC quantification and functional property assays are critical factors for cell-based therapies yet to be optimized. This study was designed to quantify the MSC population in bone marrow (BM) samples from SCD patients with osteonecrosis (SCD group) and patients with osteoarticular complications not related to SCD (NS group), using flow cytometry for CD271⁺CD45^-/low cell phenotype and CFU-F assay. We also compared expanded BM-MSC osteogenic differentiation, migration, and cytokine secretion potential between these groups. The mean total cell number, CFU-F count, and CD271⁺CD45^-/low cells in BM mononuclear concentrate were significantly higher in SCD than in NS patients. A significant correlation between CD271⁺CD45^-/low cell number and CFU-F counts was found in SCD (; ) and NS (; ) BM concentrates. An age-related quantitative reduction of CFU-F counts and CD271⁺CD45^-/low cell number was noted. Furthermore, no significant differences in the morphology, replicative capacity, expression of surface markers, multidifferentiation potential, and secretion of cytokines were found in expanded BM-MSCs from SCD and NS groups after in vitro culturing. Collectively, this work provides important data for the suitable measurement and expansion of BM-MSC in support to advanced cell-based therapies for SCD patients with osteonecrosis.

中文翻译：

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镰状细胞病伴骨坏死患者骨髓间充质基质细胞的定量和综合分析

骨髓间充质基质细胞（BM-MSC）在镰状细胞病（SCD）患者中治疗骨坏死的潜在用途正在增加。然而，方便的BM-MSC定量和功能特性测定是尚未优化的基于细胞的疗法的关键因素。这项研究旨在使用流式细胞仪检测CD271 ⁺ CD45- ^{/ low} cell来量化SCD骨坏死患者（SCD组）和骨关节并发症与SCD不相关的患者（NS组）的骨髓（BM）样本中的MSC数量。表型和CFU-F分析。我们还比较了这些组之间扩大的BM-MSC成骨分化，迁移和细胞因子分泌的潜力。平均总细胞数，CFU-F计数和CD271 ⁺ CD45SCD中BM单核浓缩液中的^{-/ low}细胞显着高于NS患者。在SCD中发现CD271 ⁺ CD45- ^/低细胞数与CFU-F计数之间存在显着相关性（; ）和NS（; ） BM精矿。注意到年龄相关的CFU-F计数和CD271 ⁺ CD45- ^/低细胞数量的减少。此外，在体外培养后，来自SCD和NS组的扩增BM-MSCs在形态，复制能力，表面标志物的表达，多分化潜能和细胞因子的分泌方面没有发现显着差异。总的来说，这项工作为适当测量和扩展BM-MSC提供了重要数据，以支持SCD骨坏死患者的先进的基于细胞的疗法。