酶解和机械解离方法对诱导多能干细胞来源的神经祖细胞的影响。
Effect of enzymatic and mechanical methods of dissociation on neural progenitor cells derived from induced pluripotent stem cells.
作者信息
Jager Lindsey D, Canda Claire-Marie A, Hall Crystal A, Heilingoetter Cassandra L, Huynh Joann, Kwok Susanna S, Kwon Jin H, Richie Jacob R, Jensen Matthew B
机构信息
Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
出版信息
Adv Med Sci. 2016 Mar;61(1):78-84. doi: 10.1016/j.advms.2015.09.005. Epub 2015 Oct 1.
PURPOSE
To determine the most effective method of dissociating neural stem and progenitor cells into a single-cell suspension.
MATERIALS/METHODS: Induced pluripotent stem cells were differentiated toward the neural fate for 4 weeks before clusters were subjected to enzymatic (Accutase, trypsin, TrypLE, dispase, or DNase I) or mechanical (trituration with pipettes of varying size) or combined dissociation. Images of cells were analyzed for cluster size using ImageJ.
RESULTS
Cells treated with the enzymes Accutase, TrypLE, or trypsin/EDTA, these enzymes followed by trituration, or a combination one of these enzymes followed by incubation with another enzyme, including DNase I, were more likely to be dissociated into a single-cell suspension.
CONCLUSIONS
Cells treated with enzymes or combinations of methods were more likely to be dissociated into a single-cell suspension.
目的
确定将神经干细胞和祖细胞解离成单细胞悬液的最有效方法。
材料/方法：诱导多能干细胞向神经命运分化4周，然后对细胞团进行酶解（Accutase、胰蛋白酶、TrypLE、Dispase或脱氧核糖核酸酶I）、机械解离（用不同大小的移液器吹打）或联合解离。使用ImageJ分析细胞图像的细胞团大小。
结果
用Accutase、TrypLE或胰蛋白酶/乙二胺四乙酸处理的细胞，这些酶处理后再进行吹打，或其中一种酶与另一种酶（包括脱氧核糖核酸酶I）联合孵育后处理的细胞，更有可能解离成单细胞悬液。
结论
用酶或方法组合处理的细胞更有可能解离成单细胞悬液。
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