The Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences Wroclaw, Wroclaw, Poland.
International Institute of Translational Medicine, Malin, Poland.
J Cell Mol Med. 2021 Jul;25(14):6634-6651. doi: 10.1111/jcmm.16667. Epub 2021 Jun 1.
Available therapies aimed at treating age-related osteoporosis are still insufficient. Therefore, designing reliable in vitro model for the analysis of molecular mechanisms underlying senile osteoporosis is highly required. We have isolated and characterized progenitor cells isolated from bone marrow (BMSCs) of osteoporotic mice strain SAM/P6 (BMSC ). The cytophysiology of BMSC was for the first time compared with BMSCs isolated from healthy BALB/c mice (BMSC ). Characterization of the cells included evaluation of their multipotency, morphology and determination of specific phenotype. Viability of BMSCs cultures was determined in reference to apoptosis profile, metabolic activity, oxidative stress, mitochondrial membrane potential and caspase activation. Additionally, expression of relevant biomarkers was determined with RT-qPCR. Obtained results indicated that BMSC and BMSC show the typical phenotype of mesenchymal stromal cells (CD44+, CD73+, CD90+) and do not express CD45. Further, BMSC were characterized by deteriorated multipotency, decreased metabolic activity and increased apoptosis occurrence, accompanied by elevated oxidative stress and mitochondria depolarisation. The transcriptome analyses showed that BMSC are distinguished by lowered expression of molecules crucial for proper osteogenesis, including Coll-1, Opg and Opn. However, the expression of Trap, DANCR1 and miR-124-3p was significantly up-regulated. Obtained results show that BMSC present features of progenitor cells with disturbed metabolism and could serve as appropriate model for in vitro investigation of age-dependent osteoporosis.
现有的针对治疗与年龄相关的骨质疏松症的疗法仍然不足。因此，设计可靠的体外模型来分析老年性骨质疏松症的分子机制是非常必要的。我们已经从骨质疏松症小鼠品系 SAM/P6 的骨髓（BMSCs）中分离并鉴定了祖细胞（BMSC）。我们首次比较了 BMSC 与来自健康 BALB/c 小鼠的 BMSCs 的细胞生理学特性。细胞特征包括评估其多能性、形态和确定特定表型。通过检测细胞凋亡谱、代谢活性、氧化应激、线粒体膜电位和半胱天冬酶激活来确定 BMSC 培养物的活力。此外，还通过 RT-qPCR 测定了相关生物标志物的表达。研究结果表明，BMSC 和 BMSC 表现出间充质基质细胞的典型表型（CD44+、CD73+、CD90+），不表达 CD45。进一步的研究表明，BMSC 的多能性恶化，代谢活性降低，凋亡发生率增加，同时伴有氧化应激和线粒体去极化增加。转录组分析表明，BMSC 中与适当成骨有关的关键分子（包括 Coll-1、Opg 和 Opn）的表达降低。然而，Trap、DANCR1 和 miR-124-3p 的表达显著上调。研究结果表明，BMSC 具有代谢紊乱的祖细胞特征，可作为体外研究年龄相关性骨质疏松症的合适模型。
