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Establishment of automated culture system for murine induced pluripotent stem cells

机译:小鼠诱导多能干细胞自动培养系统的建立

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Background Induced pluripotent stem (iPS) cells can differentiate into any cell type, which makes them an attractive resource in fields such as regenerative medicine, drug screening, or in vitro toxicology. The most important prerequisite for these industrial applications is stable supply and uniform quality of iPS cells. Variation in quality largely results from differences in handling skills between operators in laboratories. To minimize these differences, establishment of an automated iPS cell culture system is necessary. Results We developed a standardized mouse iPS cell maintenance culture, using an automated cell culture system housed in a CO2 incubator commonly used in many laboratories. The iPS cells propagated in a chamber uniquely designed for automated culture and showed specific colony morphology, as for manual culture. A cell detachment device in the system passaged iPS cells automatically by dispersing colonies to single cells. In addition, iPS cells were passaged without any change in colony morphology or expression of undifferentiated stem cell markers during the 4 weeks of automated culture. Conclusions Our results show that use of this compact, automated cell culture system facilitates stable iPS cell culture without obvious effects on iPS cell pluripotency or colony-forming ability. The feasibility of iPS cell culture automation may greatly facilitate the use of this versatile cell source for a variety of biomedical applications.
机译:背景技术诱导多能干(iPS)细胞可以分化为任何细胞类型,这使其成为再生医学,药物筛选或体外毒理学等领域的诱人资源。这些工业应用的最重要前提是iPS电池的稳定供应和统一质量。质量的差异很大程度上是由于实验室操作人员之间的处理技能差异所致。为了最大程度地减少这些差异,必须建立自动iPS细胞培养系统。结果我们使用自动细胞培养系统开发了标准化的小鼠iPS细胞维持培养,该系统安装在许多实验室常用的CO 2 培养箱中。 iPS细胞在专门为自动培养设计的培养室内繁殖,并显示出特定的菌落形态,就像人工培养一样。系统中的细胞分离装置通过将菌落分散到单个细胞中而自动传代iPS细胞。另外,在自动培养的4周中,iPS细胞在集落形态或未分化干细胞标志物的表达上没有任何变化。结论我们的结果表明,使用这种紧凑的自动化细胞培养系统有助于稳定的iPS细胞培养,而对iPS细胞的多能性或集落形成能力没有明显影响。 iPS细胞培养自动化的可行性可能会极大地促进这种多功能细胞来源在各种生物医学应用中的使用。

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