Effect of enzymatic and mechanical methods of dissociation on neural progenitor cells derived from induced pluripotent stem cells

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• 作者：Lindsey D. Jager ; Claire-Marie A. Canda ; Crystal A. Hall ; Cassandra L. Heilingoetter ; Joann Huynh ; Susanna S. Kwok ; Jin H. Kwon ; Jacob R. Richie ; Matthew B. Jensen ; ^{jensen@neurology.wisc.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Stem cells ; Neural progenitor cells ; Dissociation ; Single-cell suspension
• 刊名：Advances in Medical Sciences
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：61
• 期：1
• 页码：78-84
• 全文大小：2191 K

文摘

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.