Zhironkina O A, Shipounova I N, Bigildeev A E, Sats N V, Petinati N A, Drize N I
Hematological Research Center, Ministry of Health Care and Social Development of the Russian Federation, Moscow, Russia.
Bull Exp Biol Med. 2012 Feb;152(4):543-7. doi: 10.1007/s10517-012-1571-5.
We studied the capacity of multipotent mesenchymal stromal cells isolated from human bone marrow (BM) to long-term passaging, cloning, and re-cloning. Initial multipotent mesenchymal stromal cells and cells after gene labeling were studied. Multipotent mesenchymal stromal cells were obtained from donors (13-59 years) and cultured for 7 passages. Third generation lentivector was used for delivery of green fluorescent protein marker gene. The procedure of infection revealed reduced proliferative potential of multipotent mesenchymal stromal cells from elder donors. Hierarchy of precursor cells differing by their proliferative potential was demonstrated in the culture of multipotent mesenchymal stromal cells. Three categories of multipotent mesenchymal stromal cells were identified: mature cells incapable of proliferation (75.7±2.4% population) and cells with low and high proliferative potential (17.6±2.1 and 6.7±0.3%, respectively). The relative content of these cells insignificantly differed from passage to passage. The efficiency of cloning also remains stable, but re-cloning capacity sharply decreased after passage 3 and completely disappeared in multipotent mesenchymal stromal cells after cryopreservation. Thus, cultured multipotent mesenchymal stromal cells represent a heterogeneous and hierarchically organized population and the characteristics of this population depend of the duration of culturing and age of BM donor. This should be taken into account when using multipotent mesenchymal stromal cells in clinical practice.
我们研究了从人骨髓（BM）中分离出的多能间充质基质细胞进行长期传代、克隆和再克隆的能力。对初始多能间充质基质细胞和基因标记后的细胞进行了研究。多能间充质基质细胞取自13至59岁的供体，并培养了7代。使用第三代慢病毒载体来递送绿色荧光蛋白标记基因。感染过程显示老年供体的多能间充质基质细胞增殖潜力降低。在多能间充质基质细胞培养物中证明了具有不同增殖潜力的前体细胞层次结构。鉴定出三类多能间充质基质细胞：无增殖能力的成熟细胞（占细胞群体的75.7±2.4%）以及具有低增殖潜力和高增殖潜力的细胞（分别占17.6±2.1%和6.7±0.3%）。这些细胞的相对含量在各代之间差异不显著。克隆效率也保持稳定，但再克隆能力在第3代后急剧下降，并且在冷冻保存后的多能间充质基质细胞中完全消失。因此，培养的多能间充质基质细胞代表了一个异质性且分层组织的群体，该群体的特征取决于培养时间和骨髓供体的年龄。在临床实践中使用多能间充质基质细胞时应考虑到这一点。
