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一种新的方法,用于制备间充质干细胞并用 Ferumoxytol 进行标记,以通过 MRI 进行细胞示踪。

A New Method for Preparing Mesenchymal Stem Cells and Labeling with Ferumoxytol for Cell Tracking by MRI.

机构信息

Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, USA.

Department of Geology and Environmental Science, University of Pittsburgh, Pittsburgh, PA, USA.

出版信息

Sci Rep. 2016 May 18;6:26271. doi: 10.1038/srep26271.

Abstract

Mesenchymal stem cells (MSCs) are among the major stem cells used for cell therapy and regenerative medicine. In-vivo cell-tracking by magnetic resonance imaging (MRI) is crucial for regenerative medicine, allowing verification that the transplanted cells reach the targeted sites. Cellular MRI combined with superparamagnetic iron-oxide (SPIO) contrast agents is an effective cell-tracking method. Here, we are reporting a new "bio-mimicry" method by making use of the "in-vivo environment" of MSCs to prepare native MSCs, so that (i) the phagocytic activity of cultured MSCs can be recovered and expanded MSCs can be ex-vivo labeled with Ferumoxytol, which is currently the only FDA approved SPIO nanoparticles for human use. Using our new method, 7-day cultured MSCs regain the capability to take up Ferumoxytol and exhibit an intracellular iron concentration of 2.50 ± 0.50 pg/MSC, comparable to that obtained by using Ferumoxytol-heparin-protamine nanocomplex; and (ii) cells can be re-sized to more native size, reducing from 32.0 ± 7.2 μm to 19.5 ± 5.2 μm. Our method can be very useful for expanding MSCs and labeling with Ferumoxytol, without the need for transfection agents and/or electroporation, allowing cell-tracking by MRI in both pre-clinical and clinical studies.

摘要

间充质干细胞(MSCs)是用于细胞治疗和再生医学的主要干细胞之一。体内细胞磁共振成像(MRI)跟踪对于再生医学至关重要,它可以验证移植的细胞是否到达了目标部位。细胞 MRI 与超顺磁氧化铁(SPIO)造影剂相结合是一种有效的细胞跟踪方法。在这里,我们报告了一种新的“仿生”方法,利用 MSCs 的“体内环境”来制备天然 MSCs,从而(i)可以恢复培养的 MSCs 的吞噬活性,并可以对体外培养的 MSCs 进行 Ferumoxytol 标记,目前 Ferumoxytol 是唯一经 FDA 批准用于人体的 SPIO 纳米颗粒。使用我们的新方法,培养 7 天的 MSCs 恢复了摄取 Ferumoxytol 的能力,并表现出 2.50 ± 0.50 pg/MSC 的细胞内铁浓度,与使用 Ferumoxytol-肝素-鱼精蛋白纳米复合物获得的浓度相当;(ii)细胞可以重新调整到更接近天然的大小,从 32.0 ± 7.2 μm 减少到 19.5 ± 5.2 μm。我们的方法可以非常有用地扩展 MSCs 并进行 Ferumoxytol 标记,而无需转染剂和/或电穿孔,从而可以在临床前和临床研究中通过 MRI 进行细胞跟踪。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8be5/4870722/9731d30164ef/srep26271-f1.jpg

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