i3S-Instituto de Investigação e Inovação em Saúde, Rua Alfredo Allen, 208, 4200-135 Porto, Portugal.
INEB-Instituto de Engenharia Biomédica, Rua Alfredo Allen, 208, 4200-135 Porto, Portugal.
Molecules. 2023 Apr 13;28(8):3422. doi: 10.3390/molecules28083422.
The use of human Mesenchymal Stem Cells (hMSC) as therapeutic agents for advanced clinical therapies relies on their in vitro expansion. Over the last years, several efforts have been made to optimize hMSC culture protocols, namely by mimicking the cell physiological microenvironment, which strongly relies on signals provided by the extracellular matrix (ECM). ECM glycosaminoglycans, such as heparan-sulfate, sequester adhesive proteins and soluble growth factors at the cell membrane, orchestrating signaling pathways that control cell proliferation. Surfaces exposing the synthetic polypeptide poly(L-lysine, L-leucine) (pKL) have previously been shown to bind heparin from human plasma in a selective and concentration-dependent manner. To evaluate its effect on hMSC expansion, pKL was immobilized onto self-assembled monolayers (SAMs). The pKL-SAMs were able to bind heparin, fibronectin and other serum proteins, as demonstrated by quartz crystal microbalance with dissipation (QCM-D) studies. hMSC adhesion and proliferation were significantly increased in pKL-SAMs compared to controls, most probably related to increased heparin and fibronectin binding to pKL surfaces. This proof-of-concept study highlights the potential of pKL surfaces to improve hMSC in vitro expansion possible through selective heparin/serum protein binding at the cell-material interface.
人骨髓间充质干细胞(hMSC)作为治疗剂应用于高级临床治疗依赖于其体外扩增。在过去的几年中,已经做出了一些努力来优化 hMSC 培养方案,即通过模拟细胞生理微环境,该环境强烈依赖于细胞外基质(ECM)提供的信号。ECM 糖胺聚糖,如硫酸乙酰肝素,在细胞膜上隔离粘附蛋白和可溶性生长因子,协调控制细胞增殖的信号通路。先前已经表明,暴露于合成多肽聚(L-赖氨酸,L-亮氨酸)(pKL)的表面以选择性和浓度依赖性方式结合人血浆中的肝素。为了评估其对 hMSC 扩增的影响,将 pKL 固定到自组装单层(SAM)上。通过石英晶体微天平耗散(QCM-D)研究证明,pKL-SAMs 能够结合肝素、纤连蛋白和其他血清蛋白。与对照相比,pKL-SAMs 中的 hMSC 粘附和增殖显著增加,这很可能与 pKL 表面结合增加的肝素和纤连蛋白有关。这项概念验证研究强调了 pKL 表面通过在细胞-材料界面选择性结合肝素/血清蛋白来改善 hMSC 体外扩增的潜力。