Dergilev Konstantin V, Makarevich Pavel I, Tsokolaeva Zoya I, Boldyreva Maria A, Beloglazova Irina B, Zubkova Ekaterina S, Menshikov Mikhail Yu, Parfyonova Yelena V
Russian Cardiology Research and Production Complex, Laboratory of Angiogenesis, 121552, Moscow, Russian Federation.
Russian Cardiology Research and Production Complex, Laboratory of Angiogenesis, 121552, Moscow, Russian Federation; Lomonosov Moscow State University, Medical Research and Education Centre, Institute of Regenerative Medicine, Laboratory of gene and cell therapy, 119192, Moscow, Russian Federation.
Tissue Cell. 2017 Feb;49(1):64-71. doi: 10.1016/j.tice.2016.12.001. Epub 2016 Dec 16.
Cell sheets (CS) from c-kit+ cardiac stem cell (CSC) hold a potential for application in regenerative medicine. However, manufacture of CS may require thermoresponsive dishes, which increases cost and puts one in dependence on specific materials. Alternative approaches were established recently and we conducted a short study to compare approaches for detachment of CS from c-kit+ CSC. Our in-house developed method using chelation by Versene solution was compared to UpCell™ thermoresponsive plates in terms of CSC proliferation, viability, gap junction formation and engraftment in a model of myocardial infarction. Use of Versene solution instead of thermoresponsive dishes resulted in comparable CS thickness (approximately 100mcm), cell proliferation rate and no signs of apoptosis detected in both types of constructs. However, we observed a minor reduction of gap junction count in Versene-treated CS. At day 30 after delivery to infarcted myocardium both types of CS retained at the site of transplantation and contained comparable amounts of proliferating cells indicating engraftment. Thus, we may conclude that detachment of CS from c-kit+ CSC using Versene solution followed by mechanical treatment is an alternative to thermoresponsive plates allowing use of routinely available materials to generate constructs for cardiac repair.
来自c-kit+心脏干细胞(CSC)的细胞片(CS)在再生医学中具有应用潜力。然而,细胞片的制造可能需要热响应培养皿,这会增加成本并使人依赖特定材料。最近已建立了替代方法,我们进行了一项简短研究,比较从c-kit+ CSC分离细胞片的方法。就CSC增殖、活力、缝隙连接形成以及在心肌梗死模型中的植入情况而言,我们将内部开发的使用Versene溶液螯合的方法与UpCell™热响应板进行了比较。使用Versene溶液而非热响应培养皿产生的细胞片厚度相当(约100微米),细胞增殖率相似,并且在两种构建体中均未检测到凋亡迹象。然而,我们观察到经Versene处理的细胞片中缝隙连接数量略有减少。在植入梗死心肌30天后,两种类型的细胞片均保留在移植部位,并且含有相当数量的增殖细胞,表明已植入。因此,我们可以得出结论,使用Versene溶液然后进行机械处理从c-kit+ CSC分离细胞片是热响应板的一种替代方法,它允许使用常规可得材料来生成用于心脏修复的构建体。