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Characterization of vascular smooth muscle progenitor cells in human peripheral blood

人外周血中血管平滑肌祖细胞的表征

基本信息

批准号：
12670680
负责人：
SUGIYAMA Seigo
金额：
$ 2.24万
依托单位：
Department of Cardiovascular Medicine, Kumamoto University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670680/
关键词：
atherosaclerosis vascular smooth muscle cell differentiation restenosis blood cell fibrosis

项目摘要

Vascular smooth muscle cells (SMC) play an important role in human vascular diseases, including atherosclerosis, restenosis after angioplasty, transplant vasculopathy, and angiogenesis by producing extracellular matrix (ECM) proteins and proliferation. It has been believed that intimal SMC are migrated from media and they exhibit immature and inflammatory phenotype in human vascular lesions. Recently, we have demonstrated that circulating bone marrow-derived smooth muscle progenitor cells are involved in pathogenesis of neointimal hyperplasia in mouse aortic transplantation model, thus we gypothesized that vascular smooth muscle progenitor cells could exist in human peripheral circulation, contributing to human vascular fibroproliferative deseases. We cultured human peripheral blood mononuclear cells (PBMC) on ECM-coated plates in SMC-culture media and evaluated cellular phenotype by gene expression profiles. We first discovered expression of vascular smooth muscle α-actin (SM α-actin) … More , an essential vascular SMC marker, in circulating PBMC by RT-PCR, indicating the presence of "vascular smooth muscle progenitor cells" in human peripheral circulation. The SM α-actin positive cells were abundant in low-density PBMC faction (d<1.071 mg/mL) and the expression level of SM α-actin in PBMC varied substantially among donors. Large spindle-shaped, overlayered, SM α-actin-expressing cells ; "MC-like fibromuscular cells (FMC)" were cultured from the low-density PBMC. The SM-α-actin expression in PBMC-derived FMC increased during one month culture period. We confirmed expression of vascular SM α-actin in PBMC-dirived FMC by RT-PCR, Western blotting and immunostaining with a specific monoclonal antibody against SM α-actin (1A4). The PBMC-derived FMC had great ability of proliferation and express major SMC markers ; vimentin, SM22α, SM-1, and partially SM-2 during in vitro culture. Furthermore, they express ECM proteins, type-I and type-III collagen. These results provide a novel concept that "vascular smooth muscle progenitor cells" exist in human circulating peripheral blood and they could potentially contribute to pathogenesis of human vascular diseases and angiogenesis from luminal blood stream Less

血管平滑肌细胞(SMC)通过产生细胞外基质(ECM)蛋白和增殖，在动脉粥样硬化、血管成形术后再狭窄、移植血管病变和血管生成等人类血管疾病中发挥重要作用。已有研究认为，血管内膜SMC是从中膜中迁移出来的，在人类血管病变中表现出不成熟和炎性表型。最近，我们证实了循环中的骨髓源性平滑肌前体细胞参与了小鼠主动脉移植模型新生内膜增生的发病过程，从而推测血管平滑肌前体细胞可能存在于人外周循环中，参与了人类血管纤维增生性疾病的发生。我们将人外周血单个核细胞(PBMC)培养在SMC培养液中的ECM涂层平板上，并通过基因表达谱来评估细胞表型。首次发现血管平滑肌α-肌动蛋白(SMα-Actin)…的表达此外，一个重要的血管SMC标志物，通过RT-PCR在循环PBMC中的表达，表明在人的外周循环中存在“血管平滑肌前体细胞”。低密度(d&lt；1.071 mg/mL)外周血单核细胞中SMα-肌动蛋白阳性细胞较多，不同供者外周血单个核细胞SMα-肌动蛋白表达水平差异很大。从低密度的外周血单核细胞中培养出大的梭形、复层的SM-α-肌动蛋白表达细胞；在1个月的培养过程中，SM-α-肌动蛋白的表达增加。我们用RT-PCR法、Western blotting和抗SMα-Actin(1A4)的特异性单抗免疫染色证实了血管SMα-Actin在PBMC诱导的系膜细胞中的表达。体外培养条件下，骨髓间充质干细胞具有较强的增殖能力，并表达主要的平滑肌细胞标志物：波形蛋白、SM22α、SM-1和部分SM-2。此外，它们还表达ECM蛋白、I型和III型胶原。这些结果提供了一个新的概念，即“血管平滑肌前体细胞”存在于人类循环外周血中，它们可能在人类血管疾病的发病机制和血管新生中起重要作用。