Toosi S, Naderi-Meshkin H, Kalalinia F, Peivandi M T, Hossein Khani H, Bahrami A R, Heirani-Tabasi A, Mirahmadi M, Behravan J
Mashhad University of Medical Sciences Biotechnology Research Center, School of Pharmacy Mashhad Iran.
Academic Center for Education, Culture and Research (ACECR), Khorasan Razavi Branch Stem Cell and Regenerative Medicine Research Group Mashhad Iran.
Cell Mol Biol (Noisy-le-grand). 2016 Aug 31;62(10):68-74.
Mesenchymal stem cells (MSCs) have been considered promising tools for new clinical concepts in supporting cellular therapy and regenerative medicine. More recently, Ream/Irrigator/Aspirator (RIA) was introduced as a source of MSCs. In this study we compared MSCs derived from three different sources (iliac crest bone marrow (ICBM), adipose tissue (AT), and (RIA)) regarding the morphology, the success rate of isolating MSCs, colony frequency, expansion potential, osteogenic and chondrogenic differentiation capacity. MSCs were isolated from three different sources and flow cytometric analyses were performed for cell characterization. Colony-forming unit-fibroblast (CFU-F) assay and population doubling time (PDT) were evaluated for MSCs derived from three different sources and differentiation potential of RIA, ICBM-, and AT-MSCs were determined by staining. Additionally, gene expression profiles for tissue specific markers corresponding to osteogenesis and chondrogenesis were analyzed using real time polymerase chain reaction (RT-PCR). Cultured with the appropriate condition, osteogenic and chondrogenic differentiation could be confirmed in all MSC preparations. Flow cytometry analysis indicated that RIA- and AT-derived MSCs have more homogenous populations than ICBM-MSCs. A comparison of the colonogenic ability in different tissues by CFU-F assay after 10 days showed that more colonies are formed from RIA-MSCs than from ICBM-MSCs, and AT-MSCs. AT-MSCs, were dispersed with no obvious colonies. The RIA-MSCs underwent osteogenesis and chondrogenesis at a faster rate than ICBM and AT-MSCs. Direct comparisons of RIA- to ICBM- and AT-MSCs have shown the RIA-MSCs have higher differentiation toward osteoblast and chondrocytes compared to other sources of MSCs. Hence, RIA-MSCs may be recommended as a more suitable source for treating orthopedic disorders.
间充质干细胞（MSCs）被认为是支持细胞治疗和再生医学新临床概念的有前景的工具。最近，ream/冲洗器/吸引器（RIA）被引入作为MSCs的来源。在本研究中，我们比较了源自三种不同来源（髂嵴骨髓（ICBM）、脂肪组织（AT）和RIA）的MSCs在形态、分离MSCs的成功率、集落频率、扩增潜力、成骨和成软骨分化能力方面的差异。从三种不同来源分离MSCs，并进行流式细胞术分析以进行细胞表征。对源自三种不同来源的MSCs进行集落形成单位-成纤维细胞（CFU-F）测定和群体倍增时间（PDT）评估，并通过染色确定RIA、ICBM和AT-MSCs的分化潜力。此外，使用实时聚合酶链反应（RT-PCR）分析与成骨和软骨形成相对应的组织特异性标志物的基因表达谱。在适当条件下培养，所有MSCs制剂中均可证实成骨和成软骨分化。流式细胞术分析表明，RIA和AT来源的MSCs比ICBM-MSCs具有更均一的群体。10天后通过CFU-F测定比较不同组织中的集落形成能力，结果显示RIA-MSCs形成的集落比ICBM-MSCs和AT-MSCs更多。AT-MSCs分散，无明显集落。RIA-MSCs的成骨和成软骨分化速度比ICBM和AT-MSCs更快。RIA-MSCs与ICBM-MSCs和AT-MSCs的直接比较表明，与其他来源的MSCs相比，RIA-MSCs向成骨细胞和软骨细胞的分化更高。因此，RIA-MSCs可能被推荐为治疗骨科疾病更合适的来源。
