Le Pape Fiona, Cosnuau-Kemmat Lucie, Richard Gaëlle, Dubrana Frédéric, Férec Claude, Zal Franck, Leize Elisabeth, Delépine Pascal
Functional Genetics Department, INSERM Research Unit 1078, University of Western Brittany, European Brittany University.
Biotechnopole, HEMARINA SA, Aeropole Center, Morlaix.
Artif Organs. 2017 Apr;41(4):359-371. doi: 10.1111/aor.12892. Epub 2017 Mar 22.
Human mesenchymal stem cells (MSCs) are promising candidates for therapeutic applications such as tissue engineering. However, one of the main challenges is to improve oxygen supply to hypoxic areas to reduce oxygen gradient formation while preserving MSC differentiation potential and viability. For this purpose, a marine hemoglobin, HEMOXCell, was evaluated as an oxygen carrier for culturing human bone marrow MSCs in vitro for future three-dimensional culture applications. Impact of HEMOXCell on cell growth and viability was assessed in human platelet lysate (hPL)-supplemented media. Maintenance of MSC features, such as multipotency and expression of MSC specific markers, was further investigated by biochemical assays and flow cytometry analysis. Our experimental results highlight its oxygenator potential and indicate that an optimal concentration of 0.025 g/L HEMOXCell induces a 25%-increase of the cell growth rate, preserves MSC phenotype, and maintains MSC differentiation properties; a two-fold higher concentration induces cell detachment without altering cell viability. Our data suggest the potential interest of HEMOXCell as a natural oxygen carrier for tissue engineering applications to oxygenate hypoxic areas and to maintain cell viability, functions and "stemness." These features will be further tested within three-dimensional scaffolds.
人间充质干细胞(MSCs)是组织工程等治疗应用中很有前景的候选细胞。然而,主要挑战之一是改善向缺氧区域的氧气供应,以减少氧梯度形成,同时保持间充质干细胞的分化潜能和活力。为此,评估了一种海洋血红蛋白HEMOXCell作为氧载体,用于体外培养人骨髓间充质干细胞,以用于未来的三维培养应用。在添加人血小板裂解物(hPL)的培养基中评估了HEMOXCell对细胞生长和活力的影响。通过生化分析和流式细胞术分析进一步研究了间充质干细胞特性的维持情况,如多能性和间充质干细胞特异性标志物的表达。我们的实验结果突出了其充氧潜力,并表明0.025 g/L的最佳HEMOXCell浓度可使细胞生长速率提高25%,保持间充质干细胞表型,并维持间充质干细胞的分化特性;浓度高出两倍会导致细胞脱离,但不会改变细胞活力。我们的数据表明,HEMOXCell作为一种天然氧载体,在组织工程应用中为缺氧区域充氧并维持细胞活力、功能和“干性”具有潜在价值。这些特性将在三维支架内进一步测试。