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长骨间充质干细胞(Lb-MSCs):用于骨疾病的临床可靠细胞。

Long bone mesenchymal stem cells (Lb-MSCs): clinically reliable cells for osteo-diseases.

作者信息

Toosi Shirin, Naderi-Meshkin Hojjat, Kalalinia Fatemeh, Pievandi Mohammad Taghi, Hosseinkhani Hossein, Bahrami Ahmad Reza, Heirani-Tabasi Asieh, Mirahmadi Mahdi, Behravan Javad

机构信息

Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Stem Cell and Regenerative Medicine Research Group, Academic Center for Education, Culture and Research (ACECR), Khorasan Razavi Branch, Mashhad, Iran.

出版信息

Cell Tissue Bank. 2017 Dec;18(4):489-500. doi: 10.1007/s10561-017-9652-3. Epub 2017 Aug 16.

Abstract

Mesenchymal stem cells (MSCs) have been designated as the most reliable cells in clinics to treat osteo-diseases because of their versatile nature. MSCs, isolated from long bone (Lb-MSCs) are rarely reported and named as RIA-MSCs because of the reamer-irrigator-aspirator (RIA) device. The potential of these cells in the treatment of non-union bone fractures made them the ideal candidates to be studied for clinical practices. In this work, effect of cryopreservation on the proliferation and differentiation capabilities of long bone MSCs (Lb-MSCs) has been studied. For this purpose, Lb-MSCs were isolated via RIA device and characterized using flow cytometry and differentiation assays. Cells were cryopreserved for 3, 6 and 12 months and thereafter were characterized using differentiation assays and genetic markers specific for osteogenic, chondrogenic, and adipogenic potential quantitatively by qRT-PCR. Lb-MSCs were found expressing MSC characteristic markers defining their identity. The population doubling time (PDT) was about 2.5 ± 0.5 days and colonies appeared after 7-10 days. Differentiation potential and gene expression of 3, 6 and 12 months cryopreserved Lb-MSCs were unaltered. The results show that cryopreservation did not have an effect on the differentiation potential of human Lb-MSCs. Therefore, our work offers Lb-MSCs as clinically cells for treating osteo-diseases.

摘要

间充质干细胞(MSCs)因其多能性被认为是临床上治疗骨疾病最可靠的细胞。从长骨中分离出的间充质干细胞(Lb-MSCs)鲜有报道,由于使用了扩髓-冲洗-吸引(RIA)装置,这些细胞被命名为RIA-MSCs。这些细胞在治疗骨不连骨折方面的潜力使其成为临床实践研究的理想候选细胞。在这项研究中,研究了冷冻保存对长骨间充质干细胞(Lb-MSCs)增殖和分化能力的影响。为此,通过RIA装置分离Lb-MSCs,并使用流式细胞术和分化试验对其进行表征。将细胞冷冻保存3、6和12个月,然后使用分化试验和通过qRT-PCR定量分析成骨、软骨生成和脂肪生成潜能的特异性遗传标记物对其进行表征。发现Lb-MSCs表达定义其身份的间充质干细胞特征性标记物。群体倍增时间(PDT)约为2.5±0.5天,7-10天后出现集落。冷冻保存3、6和12个月的Lb-MSCs的分化潜能和基因表达未发生改变。结果表明,冷冻保存对人Lb-MSCs的分化潜能没有影响。因此,我们的研究为临床治疗骨疾病提供了Lb-MSCs。

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