Effective Rho-associated protein kinase inhibitor treatment to dissociate human iPS cells for suspension culture to form embryoid body-like cell aggregates
| Autor: | Koyuki Yazaki, Yoshitsugu Ohnuki, Hiroshi Kurosawa, Ayumi Horiguchi, Mami Aoyagi |
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| Rok vydání: | 2014 |
| Předmět: |
rho-Associated Kinases
Pyridines Chemistry Induced Pluripotent Stem Cells Cell Cell Culture Techniques Single application Bioengineering Embryoid body Amides Applied Microbiology and Biotechnology Suspension culture Cell dissociation Dissociation (chemistry) medicine.anatomical_structure Biochemistry Biophysics medicine Humans Induced pluripotent stem cell Protein Kinase Inhibitors Rho-associated protein kinase Embryoid Bodies Cell Aggregation Biotechnology |
| Zdroj: | Journal of Bioscience and Bioengineering. 118:588-592 |
| ISSN: | 1389-1723 |
| DOI: | 10.1016/j.jbiosc.2014.04.008 |
| Popis: | Treatment conditions using Y-27632 in the preparation of cell suspension of dissociated human pluripotent stem cells (hiPSCs) were investigated in the context of embryoid body (EB)-like cell aggregates. The effectiveness of a pretreatment with Y-27632 before cell dissociation and that of a Y-27632 treatment during cell dissociation were investigated from the viewpoint of simplicity and robustness. The duration of Y-27632 treatment in the preparation process affected the circularity and agglomeration of dissociated hiPSCs. A single application of pretreatment failed to prevent the onset of blebbing. However, a pretreatment promoted the agglomeration of dissociated hiPSCs when combined with the addition of Y-27632 to cell suspension. Our results indicate that pretreatment enhances the agglomeration potential of dissociated hiPSCs. When cell dissociation was performed in the presence of Y-27632, dissociated hiPSCs possessed the highest circularity and significant agglomerating property. It was shown that treatment with Y-27632 during cell dissociation is a simple and robust method to prepare dissociated hiPSCs for suspension culture to form EB-like cell aggregates. |
| Databáze: | OpenAIRE |
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