2D和3D培养人脐带间充质干细胞条件培养基对大鼠1型糖尿病模型具有双重作用:免疫调节和β细胞再生。

IF 5 3区 医学 Q2 IMMUNOLOGY Inflammation and Regeneration Pub Date : 2022-11-30 DOI:10.1186/s41232-022-00241-7
Basak Isildar, Serbay Ozkan, Merve Ercin, Selda Gezginci-Oktayoglu, Mahmut Oncul, Meral Koyuturk
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引用次数: 6

摘要

背景:1型糖尿病(T1D)是一种t细胞介导的自身免疫性疾病,其特征是胰岛中产生胰岛素的β细胞的不可逆破坏。辅助性和细胞毒性t细胞以及细胞因子的产生在这一过程中受损,在β细胞破坏中起协同作用,并且由于体内胰岛素缺乏而产生高血糖。间充质干细胞(MSCs)具有多能性、再生性和免疫抑制特性,是治疗自身免疫性疾病的良好工具。MSCs释放的旁分泌因子通过增加血管生成和增殖,抑制细胞凋亡,发挥免疫调节作用。在此背景下,本研究旨在研究在T1D模型中,在二维(2D)和三维(3D)环境中培养的人脐带源性间充质干细胞获得的条件培养基(CM)对MSC分泌组的治疗作用。方法:首先从人脐带分离骨髓间充质干细胞,并对其进行表征。然后,通过在二维和三维环境中培养MSCs制备两种不同的CMs。以总蛋白、IL-4、IL-10、IL-17和IFN-λ为指标分析CM的含量。在sprague - dawley型自身免疫性T1D模型大鼠中进行体内研究,在特定治疗模型框架内,腹腔注射12剂CM,持续4周。为了评估免疫调节作用,我们测定了牺牲后脾脏分离淋巴细胞中的Treg群体,并分析了血清中的IL-4、IL-10、IL-17和IFN-λ细胞因子。最后,通过标记Pdx1、Nkx6.1和胰岛素标记物来评估β细胞再生,这些标记物对β细胞的形成至关重要。结果:3D-CM的总蛋白和IL-4水平高于2D-CM。体内实验结果表明,CMs可诱导Treg种群,调节细胞因子的释放。将免疫组化结果结合评估,可以确定CM的应用显著增加了胰岛中β-细胞的比例。这种增加在3D-CM应用组中达到最高水平。结论:mscs - cm在T1D模型中具有免疫调节和β-细胞稳态/再生的双重治疗作用。此外,在制备CM的同时使用3D支架培养MSCs可以改善这种效果。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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2D and 3D cultured human umbilical cord-derived mesenchymal stem cell-conditioned medium has a dual effect in type 1 diabetes model in rats: immunomodulation and beta-cell regeneration.

Background: Type 1 diabetes (T1D) is a T-cell-mediated autoimmune disease characterized by the irreversible destruction of insulin-producing β-cells in pancreatic islets. Helper and cytotoxic T-cells and cytokine production, which is impaired by this process, take a synergetic role in β-cell destruction, and hyperglycemia develops due to insulin deficiency in the body. Mesenchymal stem cells (MSCs) appear like an excellent therapeutic tool for autoimmune diseases with pluripotent, regenerative, and immunosuppressive properties. Paracrine factors released from MSCs play a role in immunomodulation by increasing angiogenesis and proliferation and suppressing apoptosis. In this context, the study aims to investigate the therapeutic effects of MSC's secretomes by conditioned medium (CM) obtained from human umbilical cord-derived MSCs cultured in 2-dimensional (2D) and 3-dimensional (3D) environments in the T1D model.

Methods: First, MSCs were isolated from the human umbilical cord, and the cells were characterized. Then, two different CMs were prepared by culturing MSCs in 2D and 3D environments. The CM contents were analyzed in terms of total protein, IL-4, IL-10, IL-17, and IFN-λ. In vivo studies were performed in Sprague-Dawley-type rats with an autoimmune T1D model, and twelve doses of CM were administered intraperitoneally for 4 weeks within the framework of a particular treatment model. In order to evaluate immunomodulation, the Treg population was determined in lymphocytes isolated from the spleen after sacrification, and IL-4, IL-10, IL-17, and IFN-λ cytokines were analyzed in serum. Finally, β-cell regeneration was evaluated immunohistochemically by labeling Pdx1, Nkx6.1, and insulin markers, which are critical for the formation of β-cells.

Results: Total protein and IL-4 levels were higher in 3D-CM compared to 2D-CM. In vivo results showed that CMs induce the Treg population and regulate cytokine release. When the immunohistochemical results were evaluated together, it was determined that CM application significantly increased the rate of β-cells in the islets. This increase was at the highest level in the 3D-CM applied group.

Conclusion: The dual therapeutic effect of MSC-CM on immunomodulation and homeostasis/regeneration of β-cells in the T1D model has been demonstrated. Furthermore, this effect could be improved by using 3D scaffolds for culturing MSCs while preparing CM.

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来源期刊
CiteScore
11.10
自引率
1.20%
发文量
45
审稿时长
11 weeks
期刊介绍: Inflammation and Regeneration is the official journal of the Japanese Society of Inflammation and Regeneration (JSIR). This journal provides an open access forum which covers a wide range of scientific topics in the basic and clinical researches on inflammation and regenerative medicine. It also covers investigations of infectious diseases, including COVID-19 and other emerging infectious diseases, which involve the inflammatory responses. Inflammation and Regeneration publishes papers in the following categories: research article, note, rapid communication, case report, review and clinical drug evaluation.
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