在兔的外周血中检测非常小的胚胎样干细胞（VSELs）作为该疾病的早期检测工具

IF 3.7 3区医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Cytotherapy Pub Date : 2025-05-01 DOI:10.1016/j.jcyt.2025.03.041

E. Gounari , A. Komnenou , E. Kofidou , V. Karampatakis , G. Koliakos

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引用次数: 0

摘要

背景,在应激条件下动员后，在人外周血中成功检测到非常小的胚胎样干细胞（VSELs）（Bhartiya et al.2016）。本研究的目的是在药物诱导的动物模型中检测血管内皮细胞及其数量与促炎细胞因子水平和动物临床评价的相关性。方法将14只新西兰兔分为ⅰ- rvo组和ⅱ-对照组。RVO诱导组患者玻璃体内注射PD0325901，剂量为0.1mL/眼。根据我们小组最近发表的结果（Gounari et al.2022），通过临床评估、组织学检查、玻璃体液分泌细胞因子定量和视网膜组织分子分析，证实RVO诱导成功。在第0、7、14和36天收集外周血，同时按照我们之前的描述分离血管并进行计数（Gounari et al.2018）。结果VSELs计数显示，与对照组相比，第1天VSELs计数有统计学意义（*P=0.03），第2周VSELs计数明显减少，证明VSELs直接动员到外周血中。rvo诱导动物组细胞涂片H&；E染色显示，吊滴培养中具有高核质比的非常小的细胞能够形成胚体（EBs）。碱性磷酸酶染色阳性，结合Oct3/4、Nanog和Sox-2转录因子的过表达，证实了未分化细胞的存在，具有胚胎样特征，是对损伤的反应。血浆样品中TNF-α、IL-6和IL-8的分泌水平与检测到的血管内皮细胞数量有关。结论建立了一种从rvo诱导动物血液中分离和定量血管内皮细胞的方法。为了开发一种基于VSELs检测的优化方案，使其能够用于RVO或其他视网膜血管疾病的早期诊断，还需要进一步的研究。

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Detection of very small embryonic-like stem cells (VSELs) in the peripheral blood of rabbits subjected to induced retinal vein occlusion as a proposed early detection tool for the disease

Background & Aim

Very Small Embryonic-like Stem Cells (VSELs) have been successfully detected in human peripheral blood following mobilization under stressful conditions (Bhartiya et al.2016). The aim of the present study is the detection of VSELs in a pharmaceutically induced animal model and the correlation of their number with pro-inflammatory cytokine levels andanimals’ clinical evaluation.

Methodology

14 New Zealand rabbits were divided into group I-RVO and group II-control. For the RVO induction group I received intravitreal injections of PD0325901 at a dose of 0.1mL/eye. Successful RVO induction was confirmed through clinical evaluation, histological examinations, quantification of secreted cytokines in the vitreous fluid, and molecular analysis of retinal tissues according to our group's recently published results (Gounari et al.2022).

Peripheral blood was collected at days 0,7,14 and 36 while VSELs isolated and counted as we have previously described (Gounari et al.2018).

Results

Counting of VSELs showed a statistically significant increase in the number of VSELs in the first days compared to the control group (*P=0.03), which was markedly reduced after 2 weeks which proves the direct mobilization of VSELs into the peripheral blood. H&E staining of cell smears in RVO-induced animal group revealed very small sized cells with high nucleo-cytoplasmic ratio able to form embryonic bodies (EBs) in hanging-drop cultures. The positive alkaline phosphatase staining, in combination with the overexpression of the Oct3/4, Nanog and Sox-2 transcription factors confirmed the existence of undifferentiated cells with embryonic like features as a response to damage. Secreted levels of TNF-α, IL-6 and IL-8 measured in plasma samples are related to the numbers of detected-VSELs.

Conclusion

We here present a method for VSELs isolation and quantification from RVO-induced animals’ blood. Further research is needed in order to develop an optimized protocol based on VSELs detection able to be applied for the early diagnosis of RVO or other retinal vascular diseases.

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来源期刊

Cytotherapy 医学-生物工程与应用微生物

CiteScore

6.30

自引率

4.40%

发文量

683

审稿时长

49 days

期刊介绍： The journal brings readers the latest developments in the fast moving field of cellular therapy in man. This includes cell therapy for cancer, immune disorders, inherited diseases, tissue repair and regenerative medicine. The journal covers the science, translational development and treatment with variety of cell types including hematopoietic stem cells, immune cells (dendritic cells, NK, cells, T cells, antigen presenting cells) mesenchymal stromal cells, adipose cells, nerve, muscle, vascular and endothelial cells, and induced pluripotential stem cells. We also welcome manuscripts on subcellular derivatives such as exosomes. A specific focus is on translational research that brings cell therapy to the clinic. Cytotherapy publishes original papers, reviews, position papers editorials, commentaries and letters to the editor. We welcome "Protocols in Cytotherapy" bringing standard operating procedure for production specific cell types for clinical use within the reach of the readership.