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首页> 外文期刊>Biotechnology Progress >Emulsion-based encapsulation of pluripotent stem cells in hydrogel microspheres for cardiac differentiation
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Emulsion-based encapsulation of pluripotent stem cells in hydrogel microspheres for cardiac differentiation

机译：水凝胶微球中多能干细胞的基于乳液的包封，用于心脏分化

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Cardiovascular disease is the leading cause of death worldwide, and current treatments are ineffective or unavailable to majority of patients. Engineered cardiac tissue (ECT) is a promising treatment to restore function to the damaged myocardium; however, for these treatments to become a reality, tissue fabrication must be amenable to scalable production and be used in suspension culture. Here, we have developed a low-cost and scalable emulsion-based method for producing ECT microspheres from poly(ethylene glycol) (PEG)-fibrinogen encapsulated mouse embryonic stem cells (mESCs). Cell-laden microspheres were formed via water-in-oil emulsification; encapsulation occurred by suspending the cells in hydrogel precursor solution at cell densities from 5 to 60 million cells/ml, adding to mineral oil and vortexing. Microsphere diameters ranged from 30 to 570 mu m; size variability was decreased by the addition of 2% poly(ethylene glycol) diacrylate. Initial cell encapsulation density impacted the ability for mESCs to grow and differentiate, with the greatest success occurring at higher cell densities. Microspheres differentiated into dense spheroidal ECTs with spontaneous contractions occurring as early as Day 10 of cardiac differentiation; furthermore, these ECT microspheres exhibited appropriate temporal changes in gene expression and response to pharmacological stimuli. These results demonstrate the ability to use an emulsion approach to encapsulate pluripotent stem cells for use in microsphere-based cardiac differentiation.
机译：心血管疾病是全世界死亡的主要原因，目前的治疗是无效的或不可用的大多数患者。工程心脏组织（ECT）是一种有前途的治疗，以恢复损坏的心肌功能;然而，对于这些治疗成为现实，必须适用于可缩放的生产并用于悬浮培养物中的组织制造。在这里，我们开发了一种低成本和可扩展的基于乳液的方法，用于从聚（乙二醇）（PEG） - 纤维蛋白原包封的小鼠胚胎干细胞（MESCS）产生微球的基于微球。通过油内乳化形成细胞升起的微球;通过将水凝胶前体溶液中的细胞悬浮在5至60百万个细胞/ ml的细胞密度中，加入矿物油和涡旋。微球直径范围为30至570亩;通过加入2％聚（乙二醇）二丙烯酸酯，减小尺寸变异性。初始细胞封装密度影响了MESCS生长和分化的能力，在更高的细胞密度下发生了最大的成功。微球与心脏分化的第10天早期发生分为致密的球状等，随着心脏分化的第10天发生;此外，这些ECT微球表现出基因表达的适当时间变化和对药理学刺激的反应。这些结果证明了使用乳液方法来包封多能干细胞以用于微球的心脏分化。

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