Dimitriou H, Linardakis E, Martimianaki G, Stiakaki E, Perdikogianni C H, Charbord P, Kalmanti M
Department of Pediatric Hematology-Oncology, University Hospital of Heraklion University of Crete Medical School, Heraklion, Crete, Greece.
Cytotherapy. 2008;10(2):125-33. doi: 10.1080/14653240701851332.
Mesenchymal stromal cells (MSC) have become the focus of cellular therapeutics but little is known regarding bone marrow (BM) MSC derived from children. As MSC constitute part of BM stroma, we examined their properties in children with hematologic diseases.
BM MSC from children with non-malignant hematologic disorders and acute lymphoblastic leukemia (ALL) were isolated and expanded. MSC were immunophenotypically characterized and their functional characteristics were assessed by CFU-F assay and cell doubling time calculation. Their ability for trilineage differentiation was verified by molecular and histochemical methods. Apoptosis was evaluated and clonal analysis was performed.
MSC were isolated from BM of all groups. They acquired the mesenchymal-related markers from the first passage, with a simultaneous decrease of hematopoietic markers. A very low percentage of apoptotic cells was detected in all passages. The proliferative and clonogenic capacity did not differ among groups, with the exception of ALL at diagnosis, in which they were defective. Histochemical and molecular analysis of differentiated MSC yielded characteristics for adipocytes, osteoblasts and chondrocytes. Clonal analysis in a number of BM samples revealed a highly heterogeneous population of cells within each clone.
MSC from BM of children with hematologic disorders, with the exception of ALL at diagnosis, can be isolated in sufficient number and quality to serve as a potential source for clinical applications.
间充质基质细胞（MSC）已成为细胞治疗的焦点，但对于源自儿童的骨髓（BM）MSC了解甚少。由于MSC是BM基质的一部分，我们研究了其在血液系统疾病患儿中的特性。
分离并扩增非恶性血液系统疾病和急性淋巴细胞白血病（ALL）患儿的BM MSC。对MSC进行免疫表型鉴定，并通过集落形成单位 - 成纤维细胞（CFU - F）测定和细胞倍增时间计算评估其功能特性。通过分子和组织化学方法验证其向三系分化的能力。评估细胞凋亡并进行克隆分析。
所有组均从BM中分离出MSC。它们从第一代开始获得间充质相关标志物，同时造血标志物减少。在所有传代中检测到的凋亡细胞百分比非常低。除诊断时的ALL组外，各组间增殖和克隆形成能力无差异，ALL组在诊断时存在缺陷。对分化的MSC进行组织化学和分子分析得出了脂肪细胞、成骨细胞和软骨细胞的特征。对多个BM样本的克隆分析显示每个克隆内细胞群体高度异质性。
除诊断时的ALL外，血液系统疾病患儿BM中的MSC可以以足够的数量和质量分离出来，作为临床应用的潜在来源。
