Isolation of mesenchymal stromal cells (MSCs) from pretreated, hematologic patients is challenging. Especially after allogeneic hematopoietic cell transplantation (HCT), standard protocols using bone marrow aspirates fail to reliably recover sufficient cell numbers. Because MSCs are considered to contribute to processes that mainly affect the outcome after transplantation, such as an efficient lymphohematopoietic recovery, extent of graft-versus-host disease as well as the occurrence of leukemic relapse, it is of great clinical relevance to investigate MSC function in this context. Previous studies showed that MSCs can be isolated by collagenase digestion of large bone fragments of hematologically healthy patients undergoing hip replacement or knee surgeries. We have now further developed this procedure for the isolation of MSCs from hematologic patients after allogeneic HCT by using trephine biopsy specimens obtained during routine examinations. Comparison of aspirates and trephine biopsy specimens from patients after allogeneic HCT revealed a significantly higher frequency of clonogenic MSCs (colony-forming unit-fibroblast [CFU-F]) in trephine biopsy specimens (mean, 289.8 ± standard deviation 322.5 CFU-F colonies/1 × 106 total nucleated cells versus 4.2 ± 9.9; P < 0.0001). Subsequent expansion of functional MSCs isolated from trephine biopsy specimen was more robust and led to a significantly higher yield compared with control samples expanded from aspirates (median, 1.6 × 106; range, 0-2.3 × 107 P0 MSCs versus 5.4 × 104; range, 0-8.9 × 106; P < 0.0001). Using trephine biopsy specimens as MSC source facilitates the investigation of various clinical questions.

中文翻译：

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从同种异体造血细胞移植后患者的骨髓活检标本中可靠分离人间充质基质细胞

从预处理的血液病患者中分离间充质基质细胞 (MSC) 具有挑战性。特别是在同种异体造血细胞移植 (HCT) 之后，使用骨髓抽吸物的标准方案无法可靠地恢复足够的细胞数量。由于 MSC 被认为有助于影响移植后结果的过程，例如有效的淋巴造血恢复、移植物抗宿主病的程度以及白血病复发的发生，因此研究 MSC 的功能具有重要的临床意义在这种情况下。先前的研究表明，可以通过胶原酶消化接受髋关节置换术或膝关节手术的血液学健康患者的大骨碎片来分离 MSC。我们现在进一步开发了这种程序，用于通过使用在常规检查中获得的环钻活检标本在同种异体 HCT 后从血液病患者中分离 MSC。异基因 HCT 后患者的抽吸物和环钻活检标本的比较显示环钻活检标本中克隆形成的 MSC（集落形成单位-成纤维细胞 [CFU-F]）频率显着更高（平均值，289.8 ± 标准偏差 322.5 CFU-F 集落/ 1 × 106 总有核细胞对比 4.2 ± 9.9；P < 0.0001)。从环钻活检标本中分离的功能性 MSC 的后续扩增更加稳健，与从抽吸物扩增的对照样品相比，产量显着更高（中位数，1.6 × 106；范围，0-2.3 × 107 P0 MSC vs 5.4 × 104；范围， 0-8.9 × 106；P < 0.0001)。