National Institute of Science and Technology in Stem Cell and Cell Therapy, Center for Cell Therapy and Regional Blood Center, Ribeirão Preto, Brazil; Faculty of Medicine, University of São Paulo, Ribeirão Preto, Brazil.
Exp Hematol. 2010 Apr;38(4):292-300.e4. doi: 10.1016/j.exphem.2010.01.006. Epub 2010 Feb 6.
High-dose chemotherapy (HDCT) followed by autologous stem cell transplantation is a widely applied treatment for hematological and autoimmune diseases. Little is known about the effects of this therapy on multipotent mesenchymal stromal cells (MSCs). We aimed to characterize, morphologically and functionally, MSCs isolated from bone marrow aspirates of patients after HDCT.
We studied 12 consecutive lymphoma patients submitted to BEAM conditioning regimen followed by autologous stem cell transplantation 28 to 1836 days before the sample collection. Thirteen normal donors were used as control. MSCs were isolated by adherence to plastic and expanded ex vivo by culture in flasks containing alpha-minimum essential medium plus 15% fetal bovine serum.
The cell population isolated showed a typical MSC morphology, immunophenotype, and differentiation capacity into adipogenic, osteogenic, and chondrogenic lineages. The MSCs obtained from patients with Hodgkin's disease and non-Hodgkin's lymphoma showed decreased fibroblastoid colony-forming unit count (p = 0.023) and increased doubling time (p = 0.031) related to the control group. The total cell expansion of MSCs from normal subjects was marginally superior to the patient group (p = 0.064). There were no differences in gene expression profile, MSCs plasticity, or hematopoiesis support capability between control and patient group.
Results suggest that HDCT applied to lymphoma patients damaged MSCs, which was demonstrated by their reduced clonogenic potential, doubling time, and cell expansion rates when compared to controls.
大剂量化疗（HDCT）联合自体干细胞移植是一种广泛应用于血液系统和自身免疫性疾病的治疗方法。然而，对于这种治疗方法对多能间充质基质细胞（MSCs）的影响知之甚少。我们旨在对接受 HDCT 后的患者骨髓抽吸物中分离的 MSCs 进行形态学和功能特征分析。
我们研究了 12 例连续接受 BEAM 预处理方案后接受自体干细胞移植的淋巴瘤患者，采集样本的时间距移植后 28 至 1836 天。13 例正常供者作为对照。MSCs 通过贴壁法从骨髓抽吸物中分离，并在含有α-最小必需培养基和 15%胎牛血清的培养瓶中进行体外扩增。
分离得到的细胞群体表现出典型的 MSC 形态、免疫表型和向成脂、成骨和软骨谱系分化的能力。与对照组相比，来自霍奇金病和非霍奇金淋巴瘤患者的 MSCs 成纤维样集落形成单位计数减少（p=0.023），倍增时间延长（p=0.031）。正常供者的 MSC 总细胞扩增量略优于患者组（p=0.064）。对照组和患者组之间在基因表达谱、MSC 可塑性或造血支持能力方面没有差异。
结果表明，应用于淋巴瘤患者的 HDCT 损伤了 MSCs，表现为其克隆形成潜力、倍增时间和细胞扩增率降低，与对照组相比。
