Oncoematologia Pediatrica, Fondazione IRCCS Policlinico San Matteo, Università di Pavia, Pavia, Italy.
Cytotherapy. 2009;11(7):825-36. doi: 10.3109/14653240903121260.
Because of their capacity to modulate the immune response and promote tissue repair, mesenchymal stromal cells (MSC) represent a potential novel treatment for autoimmune/inflammatory diseases, including Crohn's disease (CD). The aim of the study was in vitro characterization of MSC from active CD patients for future clinical application.
MSC from the bone marrow (BM) of seven CD patients (median age 32 years) were expanded ex vivo in the presence of 5% platelet lysate; cells were investigated for clonogenic efficiency, proliferative capacity, morphology, immunophenotype, differentiation potential, genetic stability and ability to suppress in vitro proliferation of both autologous and allogeneic lymphocytes to polyclonal mitogens. Results were compared with those of BM MSC of four healthy donors (HD).
MSC were successfully expanded from all patients. Colony-forming unit-fibroblast (CFU-F) frequency and proliferative capacity were comparable in CD and HD MSC. CD MSC showed typical spindle-shaped morphology and differentiated into osteoblasts, adipocytes and chondrocytes. Surface immunologic markers did not differ between CD and HD MSC, with the only exception of sizeable levels of HLA-DR at early culture passages [12-84% at passage (P)1] in the former. CD MSC ceased their growth at variable passages (from P8 to P25) and entered senescence without any change in morphology/proliferation rate. Array-comparative genomic hybridization demonstrated that CD MSC do not show imbalanced chromosomal rearrangements. Both CD and HD MSC inhibited in vitro proliferation of lymphocytes to mitogens.
CD MSC show biologic characteristics similar to HD MSC and can be considered for anti-inflammatory and reparative cell therapy approaches in patients with refractory disease.
间充质基质细胞（MSC）因其能够调节免疫反应和促进组织修复而具有成为治疗自身免疫/炎症性疾病（包括克罗恩病（CD））的新方法的潜力。本研究的目的是对处于活动期的 CD 患者的 MSC 进行体外特征描述，以便将来进行临床应用。
从 7 名 CD 患者（中位年龄 32 岁）的骨髓（BM）中体外扩增 MSC，在存在 5%血小板裂解物的情况下进行扩增；研究细胞的集落形成效率、增殖能力、形态、免疫表型、分化潜能、遗传稳定性以及抑制自身和同种异体淋巴细胞对多克隆有丝分裂原体外增殖的能力。将结果与 4 名健康供体（HD）的 BM MSC 进行比较。
所有患者均成功扩增了 MSC。CD 和 HD MSC 的集落形成单位-成纤维细胞（CFU-F）频率和增殖能力相当。CD MSC 表现出典型的梭形形态，并分化为成骨细胞、脂肪细胞和成软骨细胞。CD 和 HD MSC 的表面免疫标志物没有差异，只有 HLA-DR 水平在早期培养阶段（第 1 代 12-84%）相当高，这是唯一的例外。CD MSC 在不同的传代（从 P8 到 P25）中停止生长并进入衰老期，形态/增殖率没有任何变化。阵列比较基因组杂交表明 CD MSC 没有不平衡的染色体重排。CD 和 HD MSC 均抑制淋巴细胞对有丝分裂原的体外增殖。
CD MSC 表现出与 HD MSC 相似的生物学特征，可考虑用于难治性疾病患者的抗炎和修复细胞治疗方法。
