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超顺磁氧化铁标记对骨髓间充质干细胞在趋化因子受体表达和趋化性中的功能的影响。

Influence of SPIO labelling on the function of BMSCs in chemokine receptors expression and chemotaxis.

机构信息

Department of Pediatrics, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou, China.

出版信息

PeerJ. 2023 Jun 2;11:e15388. doi: 10.7717/peerj.15388. eCollection 2023.

Abstract

Bone marrow-derived mesenchymal stem cells (BMSCs) are increasingly being used in bone marrow transplantation (BMT) to enable homing of the allogeneic hematopoietic stem cells and suppress acute graft versus host disease (aGVHD). The aim of this study was to optimize the labelling of BMSCs with superparamagnetic iron oxide particles (SPIOs), and evaluate the impact of the SPIOs on the biological characteristics, gene expression profile and chemotaxis function of the BMSCs. The viability and proliferation rates of the SPIO-labeled BMSCs were analyzed by trypan blue staining and CCK-8 assay respectively, and the chemotaxis function was evaluated by the transwell assay. The expression levels of chemokine receptors were measured by RT-PCR and flow cytometry. The SPIOs had no effect on the viability of the BMSCs regardless of the labelling concentration and culture duration. The labelling rate of the cells was higher when cultured for 48 h with the SPIOs. Furthermore, cells labeled with 25 µg/ml SPIOs for 48 h had the highest proliferation rates, along with increased expression of chemokine receptor genes and proteins. However, there was no significant difference between the chemotaxis function of the labeled and unlabeled BMSCs. To summarize, labelling BMSCs with 25 µg/ml SPIOs for 48h did not affect their biological characteristics and chemotaxis function, which can be of significance for applications.

摘要

骨髓间充质干细胞(BMSCs)越来越多地被用于骨髓移植(BMT)中,以使同种异体造血干细胞归巢,并抑制急性移植物抗宿主病(aGVHD)。本研究旨在优化超顺磁性氧化铁颗粒(SPIOs)对 BMSCs 的标记,并评估 SPIOs 对 BMSCs 生物学特性、基因表达谱和趋化功能的影响。通过台盼蓝染色和 CCK-8 检测分别分析 SPIO 标记的 BMSCs 的活力和增殖率,通过 Transwell 检测评估趋化功能。通过 RT-PCR 和流式细胞术测量趋化因子受体的表达水平。无论标记浓度和培养时间如何,SPIOs 对 BMSCs 的活力均无影响。当用 SPIOs 培养 48 小时时,细胞的标记率更高。此外,用 25μg/ml SPIOs 标记 48 小时的细胞具有最高的增殖率,同时趋化因子受体基因和蛋白的表达增加。然而,标记和未标记的 BMSCs 的趋化功能没有显著差异。总之,用 25μg/ml SPIOs 标记 BMSCs 48 小时不会影响其生物学特性和趋化功能,这对于应用具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72f0/10241165/a21c86c3b534/peerj-11-15388-g001.jpg

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