Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Korea.
BMC Vet Res. 2012 Aug 27;8:145. doi: 10.1186/1746-6148-8-145.
Development of a method for long-term labeling of cells is critical to elucidate transplanted cell fate and migration as well as the contribution to tissue regeneration. Silica nanoparticles have been recently developed and demonstrated to be biocompatible with a high labeling capacity. Thus, our study was designed to assess the suitability of silica nanoparticles for labeling canine mesenchymal stem cells (MSCs) and the fluorescence afficiency in highly autofluorescent tissue.
We examined the effect of silica nanoparticle labeling on stem cell morphology, viability and differentiation as compared with those of unlabeled control cells. After 4 h of incubation with silica nanoparticles, they were internalized by canine MSCs without a change in the morphology of cells compared with that of control cells. The viability and proliferation of MSCs labeled with silica nanoparticles were evaluated by a WST-1 assay and trypan blue exclusion. No effects on cell viability were observed, and the proliferation of canine MSCs was not inhibited during culture with silica nanoparticles. Furthermore, adipogenic and osteogenic differentiation of silica nanoparticle-labeled canine MSCs was at a similar level compared with that of unlabeled cells, indicating that silica nanoparticle labeling did not alter the differentiation capacity of canine MSCs. Silica nanoparticle-labeled canine MSCs were injected into the kidneys of BALB/c mice after celiotomy, and then the mice were sacrificed after 2 or 3 weeks. The localization of injected MSCs was closely examined in highly autofluorescent renal tissues. Histologically, canine MSCs were uniformly and completely labeled with silica nanoparticles, and were unambiguously imaged in histological sections.
The results of the current study showed that silica nanoparticles are useful as an effective labeling marker for MSCs, which can elucidate the distribution and fate of transplanted MSCs.
开发一种长期标记细胞的方法对于阐明移植细胞的命运和迁移以及对组织再生的贡献至关重要。硅纳米粒子最近已被开发出来,并被证明具有生物相容性和高标记能力。因此,我们的研究旨在评估硅纳米粒子标记犬间充质干细胞(MSCs)的适用性以及在高度自发荧光组织中的荧光效率。
我们研究了与未标记对照细胞相比,硅纳米粒子标记对干细胞形态、活力和分化的影响。用硅纳米粒子孵育 4 小时后,它们被犬 MSCs 内化,与对照细胞相比,细胞形态没有变化。通过 WST-1 测定和台盼蓝排斥试验评估了用硅纳米粒子标记的 MSC 的活力和增殖。未观察到对细胞活力的影响,并且在含有硅纳米粒子的培养过程中,犬 MSCs 的增殖未受到抑制。此外,与未标记细胞相比,硅纳米粒子标记的犬 MSCs 的成脂和成骨分化水平相似,表明硅纳米粒子标记未改变犬 MSCs 的分化能力。在剖腹手术后,将硅纳米粒子标记的犬 MSCs 注入 BALB/c 小鼠肾脏,然后在 2 或 3 周后处死小鼠。在高度自发荧光的肾脏组织中密切检查注射 MSCs 的定位。组织学上,犬 MSCs 被硅纳米粒子均匀且完全标记,并且在组织学切片中可以明确成像。
本研究结果表明,硅纳米粒子可用作 MSC 的有效标记物,可阐明移植 MSC 的分布和命运。