纤维蛋白对非限制性体细胞干细胞粘附分子（ICAM-1、ITGAV和ITGB3）表达的影响

IF 1.8 Q3 HEMATOLOGY

Hematology, Transfusion and Cell Therapy Pub Date : 2025-04-01 DOI:10.1016/j.htct.2025.103827

Sanaz Khaseb , Mahdi Kohansal Vajari , Mina Soufi Zomorrod , Maryam Rezai Rad , Monireh Ajami , Mansoureh Ajami , Saba Sadeghpour , Amir Atashi

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引用次数: 0

摘要

造血干细胞的扩增依赖于粘附分子、整合素和细胞因子介导的直接细胞间相互作用。不受限制的体细胞干细胞作为一种新的基质细胞，通过分泌造血相关细胞因子和表达粘附分子，在共培养条件下支持造血干细胞的扩增。先前的研究表明，纤维蛋白增加了这些细胞中造血相关基因的表达。本研究主要研究非限制性体细胞干细胞在三维纤维蛋白支架上的粘附特性。方法从脐带血中分离非限制性体细胞干细胞，采用流式细胞术和多系分化技术对其进行鉴定。扫描电镜和DAPI染色分析细胞与纤维蛋白的附着。MTT法测定纤维蛋白活性。采用定量聚合酶链反应评价三维纤维蛋白支架培养细胞中细胞间粘附分子1 （ICAM-1）、整合素亚基αv （ITGAV）、整合素亚基β3 （ITGB3）的表达。结果CD73、CD105、CD166表达阳性，CD45表达阴性。茜素红和油红O染色证实钙沉积和脂质液泡。MTT试验显示纤维蛋白对细胞活力有积极影响。与在塑料组织培养板上培养的细胞相比，在纤维蛋白上培养的细胞中ITGAV的表达显著增加（对照组）。此外，ITGB3的表达在两组中均无明显变化，而ICAM-1的表达在纤维蛋白培养的细胞中下调。结论研究表明，纤维蛋白对ITGAV的表达有正向影响，而ITGAV在造血干细胞增殖的直接细胞间相互作用中起重要作用。

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Effect of fibrin on the expression of adhesion molecules (ICAM-1, ITGAV, and ITGB3) in unrestricted somatic stem cells

Background

Hematopoietic stem cell expansion relies on direct cell-cell interactions mediated by adhesion molecules, integrins, and cytokines. Unrestricted somatic stem cells have emerged as novel stromal cells supporting hematopoietic stem cell expansion in co-culture conditions via secretion of hematopoiesis-related cytokines and the expression of adhesion molecules. Previous research showed fibrin increased hematopoiesis-related gene expression in these cells. This study focused on the adhesive characteristics of unrestricted somatic stem cells on 3D fibrin scaffolds.

Methods

Unrestricted somatic stem cells were isolated from umbilical cord blood and characterized using flow cytometry and multilineage differentiation assays. Scanning electron microscopy and DAPI staining were employed to analyze cell attachment to fibrin. Viability on fibrin was assessed through MTT assays. Quantitative polymerase chain reaction was conducted to evaluate the expression of intercellular adhesion molecule 1 (ICAM-1), integrin subunit αv (ITGAV), and integrin subunit β3 (ITGB3) in cells cultured on 3D fibrin scaffolds.

Results

Cells were positive for CD73, CD105, and CD166 but negative for CD45. Alizarin red and Oil red O stains confirmed calcium deposition and lipid vacuoles. MTT assays revealed that fibrin positively impacts viability. ITGAV expression was significantly increased in cells cultured on fibrin compared to those cultured on plastic tissue culture plates (Control Group). Furthermore, ITGB3 expression showed no significant change in both groups, while ICAM-1 expression was downregulated in cells cultured on fibrin.

Conclusions

Our study revealed that fibrin has a positive impact on the expression of ITGAV, which plays a crucial role in direct cell-cell interactions affecting hematopoietic stem cell expansion.

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