免疫抑制药物对从糖尿病和外周动脉疾病患者中分离出的骨髓源性干细胞的特性和功能特性的影响。

The Effects of Immunosuppressive Drugs on the Characteristics and Functional Properties of Bone Marrow-Derived Stem Cells Isolated from Patients with Diabetes Mellitus and Peripheral Arterial Disease.

作者信息

Husakova Jitka, Echalar Barbora, Kossl Jan, Palacka Katerina, Fejfarova Vladimira, Dubsky Michal

机构信息

Diabetes Centre, Institute for Clinical and Experimental Medicine, 14021 Prague, Czech Republic.

First Faculty of Medicine, Charles University, 14021 Prague, Czech Republic.

出版信息

Biomedicines. 2023 Jun 30;11(7):1872. doi: 10.3390/biomedicines11071872.

Abstract

BACKGROUND

Diabetic patients (DPs) with foot ulcers can receive autologous cell therapy (ACT) as a last therapeutic option. Even DPs who have undergone organ transplantation and are using immunosuppressive (IS) drugs can be treated by ACT. The aim of our study was to analyze the effects of IS drugs on the characteristics of bone marrow-derived stem cells (BM-MSCs).

METHODS

The cells were isolated from the bone marrow of DPs, cultivated for 14-18 days, and phenotypically characterized using flow cytometry. These precursor cells were cultured in the presence of various IS drugs. The impact of IS drugs on metabolic activity was measured using a WST-1 assay, and the expression of genes for immunoregulatory molecules was detected through RT-PCR. Cell death was analyzed through the use of flow cytometry, and the production of cytokines was determined by ELISA.

RESULTS

The mononuclear fraction of cultured cells contained mesenchymal stem cells (CD45CD73CD90CD105), myeloid angiogenic cells (CD45CD146), and endothelial colony-forming cells (CD45CD146). IS drugs inhibited metabolic activity, the expression of genes for immunoregulatory molecules, the production of cytokines, and the viability of the cells.

CONCLUSIONS

The results indicate that IS drugs in a dose-dependent manner had a negative impact on the properties of BM-MSCs used to treat ischemic diabetic foot ulcers, and that these drugs could affect the therapeutic potential of BM-MSCs.

摘要

背景

患有足部溃疡的糖尿病患者(DPs)可将自体细胞疗法(ACT)作为最后的治疗选择。即使是接受过器官移植并正在使用免疫抑制(IS)药物的DPs也可接受ACT治疗。我们研究的目的是分析IS药物对骨髓来源干细胞(BM-MSCs)特性的影响。

方法

从DPs的骨髓中分离细胞,培养14 - 18天,并使用流式细胞术进行表型鉴定。这些前体细胞在各种IS药物存在的情况下进行培养。使用WST-1检测法测量IS药物对代谢活性的影响,并通过RT-PCR检测免疫调节分子基因的表达。通过流式细胞术分析细胞死亡情况,并通过ELISA测定细胞因子的产生。

结果

培养细胞的单核部分包含间充质干细胞(CD45CD73CD90CD105)、髓系血管生成细胞(CD45CD146)和内皮集落形成细胞(CD45CD146)。IS药物抑制了代谢活性、免疫调节分子基因的表达、细胞因子的产生以及细胞的活力。

结论

结果表明,IS药物以剂量依赖的方式对用于治疗缺血性糖尿病足溃疡的BM-MSCs的特性产生负面影响,并且这些药物可能会影响BM-MSCs的治疗潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9626/10377428/adb6733bad8d/biomedicines-11-01872-g001.jpg

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