冷冻和储存对分离用于临床的人间充质基质细胞特性和功能的影响。
The influence of freezing and storage on the characteristics and functions of human mesenchymal stromal cells isolated for clinical use.
作者信息
Haack-Sorensen M, Bindslev L, Mortensen S, Friis T, Kastrup J
机构信息
Cardiology Stem Cell Laboratory, University Hospital Rigshospitalet, Copenhagen, Denmark.
出版信息
Cytotherapy. 2007;9(4):328-37. doi: 10.1080/14653240701322235.
BACKGROUND
Studies have shown that stem cell therapy could be a novel option for improving neovascularization and cardiac function in patients with ischemic heart disease. Human mesenchymal stromal cells (MSC) have generated wide interest in the clinical setting because of their ability to regenerate tissue. The aim of the study was to test whether freezing and storage of human BM mononuclear cells (BM-MNC) and ex vivo-expanded MSC influenced their phenotypic and functional characteristics as well as proliferation capacity.
METHODS
MNC were isolated from BM and divided into two portions: one part was immediately cultured (MSC P0) whereas the second part was frozen for a week before cultivation and analysis (F-MSC P1). Confluent MSC (P0) were harvested and divided: one was analyzed as MSC P1 and the other was frozen for a week before further cultivation and analysis as F-MSC P2.
RESULTS
MSC P1, F-MSC P1 and F-MSC P2 had similar proliferation capacities and demonstrated almost identical expression levels of markers characteristic for MSC. The capacity to form endothelial vascular structures was independent of freezing.
DISCUSSION
The proliferation and differentiation capacity as well as the cellular characteristics were identical in cultivated MSC derived from freshly isolated BM-MNC and MSC derived after freezing and storage of either freshly isolated BM-MNC or ex vivo-cultivated MSC. This highlights the potential clinical use of MSC in patients with cardiac and degenerative diseases, as it would be possible to inject MSC obtained from the same BM aspiration at different time points.
背景
研究表明,干细胞疗法可能是改善缺血性心脏病患者新血管形成和心脏功能的一种新选择。人间充质基质细胞(MSC)因其组织再生能力而在临床环境中引起了广泛关注。本研究的目的是测试人骨髓单个核细胞(BM-MNC)的冷冻保存以及体外扩增的MSC是否会影响其表型、功能特性以及增殖能力。
方法
从骨髓中分离单个核细胞并分为两部分:一部分立即进行培养(MSC P0),而另一部分在培养和分析前冷冻一周(F-MSC P1)。收获汇合的MSC(P0)并进行划分:一部分作为MSC P1进行分析,另一部分在进一步培养和分析前冷冻一周作为F-MSC P2。
结果
MSC P1、F-MSC P1和F-MSC P2具有相似的增殖能力,并显示出几乎相同的MSC特征性标志物表达水平。形成内皮血管结构的能力与冷冻无关。
讨论
源自新鲜分离的BM-MNC的培养MSC以及源自新鲜分离的BM-MNC或体外培养的MSC冷冻保存后的MSC,其增殖和分化能力以及细胞特征是相同的。这突出了MSC在心脏和退行性疾病患者中的潜在临床应用,因为有可能在不同时间点注射从同一骨髓穿刺获得的MSC。