Tangchitphisut Paween, Srikaew Narongrit, Phongkitkarun Sith, Jaovisidha Suphaneewan, Tawonsawatruk Tulyapruek
Department of Orthopaedics, School of Medicine, Mae Fah Luang University, Thailand.
Research and Innovative Center, Ramathibodi Hospital, Thailand.
Heliyon. 2020 Aug 1;6(8):e04582. doi: 10.1016/j.heliyon.2020.e04582. eCollection 2020 Aug.
The objective of this study was to investigate iron sucrose labeling in mesenchymal stem cell (MSCs) tracking.
Adipose-derived mesenchymal stem cell-based therapy is a promising strategy for promoting musculoskeletal repair.
Iron sucrose-labeled adipose-derived mesenchymal stem cells (IS-labeled ASCs) were tracked using T2-and T2∗-weighted sequences by 1.5 and 3 T MRI in an model. ASCs were isolated from cosmetic liposuction specimens. ASCs from passages 4-6 were labeled with iron sucrose (Venofer®) which was added to the cell culture medium. Pre- and post-iron sucrose labeled ASCs were evaluated for cell surface immunophenotypes. Cell viability as well as chondrogenic, adipogenic and osteogenic differentiation of IS-labeled-ASCs were evaluated. The IS-labeled ASCs were titrated into microtubes at 1 × 10, 1 × 10, 1 × 10 and 1 × 10 cells/ml/microtube and their intensities were determined by 1.5 and 3T MRI using T2-and T2∗-weighted sequences.
The expression markers of IS-labeled ASCs from flow cytometry were equivalent to control. The mean cell viability was 97.73 ± 2.06%. Cell differentiations of IS-labeled ASCs were confirmed in each lineage using specific staining solutions. T2∗-weighted sequences (T2∗) were able to detect iron sucrose labeled-ASCs at a minimum of 1 × 10 cells/ml/microtube using 1.5 and 3T MRI, but the detection sensitivity was lower with T2-weighted sequences (T2).
Iron sucrose incubation is a safe alternative method for ASCs labeling and tracking using MRI following treatment. Clinicians and researchers should be able to visualize the location of ASCs engraftment without secondary surgical investigation involving tissue sampling.
本研究的目的是调查铁蔗糖标记在间充质干细胞(MSC)追踪中的应用。
基于脂肪来源间充质干细胞的治疗是促进肌肉骨骼修复的一种有前景的策略。
在一个模型中,使用1.5和3T磁共振成像(MRI)的T2加权和T2加权序列追踪铁蔗糖标记的脂肪来源间充质干细胞(IS标记的ASC)。从美容抽脂标本中分离出ASC。第4至6代的ASC用添加到细胞培养基中的铁蔗糖(Venofer®)进行标记。对铁蔗糖标记前后的ASC进行细胞表面免疫表型评估。评估IS标记的ASC的细胞活力以及软骨生成、脂肪生成和成骨分化。将IS标记的ASC以1×10、1×10、1×10和1×10个细胞/毫升/微管的浓度滴定到微管中,并使用1.5和3T MRI的T2加权和T2加权序列测定其强度。
流式细胞术检测的IS标记的ASC的表达标志物与对照相当。平均细胞活力为97.73±2.06%。使用特异性染色溶液在每个谱系中证实了IS标记的ASC的细胞分化。T2加权序列(T2)能够使用1.5和3T MRI在最低1×10个细胞/毫升/微管的浓度下检测到铁蔗糖标记的ASC,但T2加权序列(T2)的检测灵敏度较低。
铁蔗糖孵育是治疗后使用MRI对ASC进行标记和追踪的一种安全替代方法。临床医生和研究人员应该能够在不进行涉及组织采样的二次手术调查的情况下可视化ASC植入的位置。