Labarre Klaus Werner, Grathwol Peter Ansgar, Zimmermann Gerald
Department of Trauma Surgery, Theresienkrankenhaus, Bassermannstraße 1, 68165 Mannheim, Germany.
Bone Rep. 2024 Nov 26;24:101820. doi: 10.1016/j.bonr.2024.101820. eCollection 2025 Mar.
Mesenchymal stem cells (MSCs) derived from bone marrow play an increasingly important role in regenerative medicine due to their capacity to promote tissue regeneration in various clinical contexts. Applications include the treatment of osteoarthritis, bone regeneration post-injury, and the management of conditions such as Crohn's disease, alopecia, and nervous system reconstruction. Accurate quantification of MSCs within Bone Marrow Concentrates (BMCs) is essential for ensuring the quality and efficacy of these cell therapy products in clinical settings.
This study aims to quantify the population of CD271 and CD45 cells in BMCs prepared using the method we have selected and to provide a basis for comparing these results with other BMC products. Additionally, we seek to determine whether the total cell count in BMCs can serve as a reliable indicator of MSC numbers and if cellularity (the number of cells per ml) can predict a higher percentage of MSCs within the population.
Bone Marrow Aspirates (BMA) were collected from 41 patients undergoing knee or hip arthroplasty. Aspirates were processed using density gradient centrifugation and positive selection of CD271 cells. Flow cytometry was applied to analyze cell subsets, and cell counts were determined with a NucleoCounter. The relationships between BMA cellularity (total cells per ml), MSC concentration (MSC count per ml), and MSC percentage (the proportion of MSCs within the total cell population) were assessed.
The mean percentage of CD271 CD45 cells in bone marrow samples was 0.03 % (SD 0.03 %). Cellularity varied significantly among samples, with a mean of 6 million cells/ml (SD 8.7 million cells/ml). A strong correlation was observed between BMC cellularity and MSC concentration ( < 0.05), although no correlation was found between cellularity and the MSC percentage.
Despite high variability in cellularity, the concentration of MSCs correlated strongly with BMC cellularity, suggesting that total cell counts can be used to estimate MSC numbers in BMCs. However, cellularity is not an indicator of a particularly high MSC content. This study supports the use of cell counts as a measure for estimating MSC concentration in BMCs. Future research should focus on establishing direct comparisons with other BMC products and exploring factors influencing cellularity and MSC percentages to enhance BMC quality for clinical applications.
源自骨髓的间充质干细胞（MSCs）因其在各种临床环境中促进组织再生的能力，在再生医学中发挥着越来越重要的作用。其应用包括骨关节炎的治疗、损伤后骨再生以及克罗恩病、脱发和神经系统重建等病症的管理。准确量化骨髓浓缩物（BMCs）中的间充质干细胞对于确保这些细胞治疗产品在临床环境中的质量和疗效至关重要。
本研究旨在量化使用我们所选方法制备的BMCs中CD271和CD45细胞的数量，并为将这些结果与其他BMC产品进行比较提供依据。此外，我们试图确定BMCs中的总细胞计数是否可作为间充质干细胞数量的可靠指标，以及细胞密度（每毫升细胞数）是否能够预测群体中间充质干细胞的更高百分比。
从41例接受膝关节或髋关节置换术的患者中采集骨髓抽吸物（BMA）。使用密度梯度离心法和CD271细胞阳性选择法对抽吸物进行处理。应用流式细胞术分析细胞亚群，并用NucleoCounter测定细胞计数。评估BMA细胞密度（每毫升总细胞数）、间充质干细胞浓度（每毫升间充质干细胞计数）和间充质干细胞百分比（间充质干细胞在总细胞群体中的比例）之间关系。
骨髓样本中CD271 CD45细胞的平均百分比为0.03%（标准差0.03%）。样本间细胞密度差异显著，平均值为600万个细胞/毫升（标准差870万个细胞/毫升）。观察到BMC细胞密度与间充质干细胞浓度之间存在强相关性（<0.05），尽管未发现细胞密度与间充质干细胞百分比之间存在相关性。
尽管细胞密度存在高度变异性，但间充质干细胞浓度与BMC细胞密度密切相关，这表明总细胞计数可用于估计BMCs中间充质干细胞数量。然而，细胞密度并非间充质干细胞含量特别高的指标。本研究支持使用细胞计数作为估计BMCs中间充质干细胞浓度的一种方法。未来研究应侧重于与其他BMC产品建立直接比较，并探索影响细胞密度和间充质干细胞百分比的因素，以提高BMCs在临床应用中的质量。
