Razban Vahid, Lotfi Abbas Sahebqadam, Soleimani Masoud, Ahmadi Hossein, Massumi Mohammad, Khajeh Sahar, Ghaedi Mahboobeh, Arjmand Sareh, Najavand Saeed, Khoshdel Alireza
National Institute of Genetic Engineering and Biotechnology (NIGEB) , Tehran, Iran .
Biores Open Access. 2012 Aug;1(4):174-83. doi: 10.1089/biores.2012.9905.
Stem cell therapy continues to be an innovative and promising strategy for heart failure. Stem cell injection alone, however, is hampered by poor cell survival and differentiation. This study was aimed to explore the possibility of improving stem cell therapy through genetic modification of stem cells, in order for them to promote angiogenesis in an auto- and paracrine manner under hypoxic conditions. Hypoxia inducible factor-1α was overexpressed in bone marrow-derived mesenchymal stem cells (MSCs) by stable transduction using a lentiviral vector. Under hypoxic and normoxic conditions, the vascular endothelial growth factor (VEGF) concentration in the cells' supernatant was measured by an enzyme-linked immunosorbent assay. Migration was assayed by wound healing and c-Met expression by flow cytometry. Tube formation was evaluated on a Matrigel basement membrane. The concentration of VEGF was significantly increased in the supernatant of HIF-1α-overexpressing MSCs; this medium was significantly more effective in inducing endothelial cell migration compared to untransduced MSCs. Transduced cells showed increased levels of c-Met expression and were more efficient at tube formation. However, no indication of differentiation toward an endothelial phenotype was observed. This study indicated that genetic modification of MSCs by HIF-1α overexpression has the potential to improve components of the angiogenesis process under a hypoxic condition by paracrine and autocrine mechanisms.
干细胞疗法仍然是治疗心力衰竭的一种创新且有前景的策略。然而，单纯的干细胞注射受到细胞存活率低和分化不良的阻碍。本研究旨在探索通过对干细胞进行基因改造来改善干细胞疗法的可能性，以便它们在缺氧条件下以自分泌和旁分泌方式促进血管生成。通过使用慢病毒载体进行稳定转导，使缺氧诱导因子-1α在骨髓间充质干细胞（MSCs）中过表达。在缺氧和常氧条件下，通过酶联免疫吸附测定法测量细胞上清液中血管内皮生长因子（VEGF）的浓度。通过伤口愈合试验检测迁移能力，通过流式细胞术检测c-Met表达。在基质胶基底膜上评估管腔形成。在过表达HIF-1α的MSCs的上清液中，VEGF的浓度显著增加；与未转导的MSCs相比，这种培养基在诱导内皮细胞迁移方面明显更有效。转导的细胞显示出c-Met表达水平增加，并且在管腔形成方面更有效。然而，未观察到向内皮表型分化的迹象。本研究表明，通过过表达HIF-1α对MSCs进行基因改造有可能通过旁分泌和自分泌机制在缺氧条件下改善血管生成过程的各个组成部分。
