Department of Natural Sciences, Bonn-Rhein-Sieg University of Applied Sciences, D-53359 Rheinbach, Germany.
Institute of Pharmaceutical & Medicinal Chemistry, University of Dusseldorf, D-40225 Dusseldorf, Germany.
Int J Mol Sci. 2020 Aug 27;21(17):6210. doi: 10.3390/ijms21176210.
Human mesenchymal stem cells (hMSCs) have shown their multipotential including differentiating towards endothelial and smooth muscle cell lineages, which triggers a new interest for using hMSCs as a putative source for cardiovascular regenerative medicine. Our recent publication has shown for the first time that purinergic 2 receptors are key players during hMSC differentiation towards adipocytes and osteoblasts. Purinergic 2 receptors play an important role in cardiovascular function when they bind to extracellular nucleotides. In this study, the possible functional role of purinergic 2 receptors during MSC endothelial and smooth muscle differentiation was investigated.
Human MSCs were isolated from liposuction materials. Then, endothelial and smooth muscle-like cells were differentiated and characterized by specific markers via Reverse Transcriptase-PCR (RT-PCR), Western blot and immunochemical stainings. Interestingly, some purinergic 2 receptor subtypes were found to be differently regulated during these specific lineage commitments: P2Y4 and P2Y14 were involved in the early stage commitment while P2Y1 was the key player in controlling MSC differentiation towards either endothelial or smooth muscle cells. The administration of natural and artificial purinergic 2 receptor agonists and antagonists had a direct influence on these differentiations. Moreover, a feedback loop via exogenous extracellular nucleotides on these particular differentiations was shown by apyrase digest.
Purinergic 2 receptors play a crucial role during the differentiation towards endothelial and smooth muscle cell lineages. Some highly selective and potent artificial purinergic 2 ligands can control hMSC differentiation, which might improve the use of adult stem cells in cardiovascular tissue engineering in the future.
人类间充质干细胞（hMSCs）具有多能性，包括向内皮细胞和平滑肌细胞谱系分化，这引发了使用 hMSCs 作为心血管再生医学潜在来源的新兴趣。我们最近的出版物首次表明，嘌呤能 2 受体是 hMSC 向脂肪细胞和成骨细胞分化过程中的关键参与者。嘌呤能 2 受体在与细胞外核苷酸结合时在心血管功能中发挥重要作用。在这项研究中，研究了嘌呤能 2 受体在 MSC 内皮和平滑肌分化过程中的可能功能作用。
从吸脂材料中分离出人 MSC。然后，通过逆转录聚合酶链反应（RT-PCR）、Western blot 和免疫化学染色，用特定标志物对内皮样细胞和平滑肌样细胞进行分化和鉴定。有趣的是，在这些特定谱系承诺期间，发现一些嘌呤能 2 受体亚型被不同程度地调节：P2Y4 和 P2Y14 参与早期承诺，而 P2Y1 是控制 MSC 向内皮或平滑肌细胞分化的关键因素。天然和人工嘌呤能 2 受体激动剂和拮抗剂的给药对这些分化有直接影响。此外，通过外源性细胞外核苷酸的无酶消化显示了这些特定分化的反馈回路。
嘌呤能 2 受体在向内皮细胞和平滑肌细胞谱系分化过程中起着至关重要的作用。一些高度选择性和有效的人工嘌呤能 2 配体可以控制 hMSC 分化，这可能会改善未来在心血管组织工程中使用成人干细胞。
