論文 - 詳細
| RRC ID | 52194 |
|---|---|
| 著者 | Kondo Y, Hattori K, Tashiro S, Nakatani E, Yoshimitsu R, Satoh T, Sugiura S, Kanamori T, Ohnuma K. |
| タイトル | Compartmentalized microfluidic perfusion system to culture human induced pluripotent stem cell aggregates. |
| ジャーナル | J Biosci Bioeng |
| Abstract |
Microfluidic perfusion systems enable small-volume cell cultures under precisely controlled microenvironments, and are typically developed for cell-based high-throughput screening. However, most such systems are designed to manipulate dissociated single cells, not cell aggregates, and are thus unsuitable to induce differentiation in human induced pluripotent stem cells (hiPSCs), which is conventionally achieved by using cell aggregates to increase cell-cell interactions. We have now developed a compartmentalized microfluidic perfusion system with large flow channels to load, culture, and observe cell aggregates. Homogeneously sized cell aggregates to be loaded into the device were prepared by shredding flat hiPSC colonies into squares. These aggregates were then seeded into microchambers coated with fibronectin and bovine serum albumin (BSA) to establish adherent and floating cultures, respectively, both of which are frequently used to differentiate hiPSCs. However, the number of aggregates loaded in fibronectin-coated microchambers was much lower than in BSA-coated microchambers, suggesting that fibronectin traps cell aggregates before they reach the chambers. Accordingly, hiPSCs that reached the microchambers subsequently adhered. In contrast, BSA-coated microchambers did not allow cell aggregates to adhere, but were sufficiently deep to prevent cell aggregates from flowing out during perfusion of media. Immunostaining for markers of undifferentiated cells showed that cultures on both fibronectin- and BSA-coated microchambers were successfully established. Notably, we found that floating aggregates eventually adhered to surfaces coated with BSA upon differentiation, and that differentiation depends on the initial size of aggregates. Collectively, these results suggest that the microfluidic system is suitable for manipulating hiPSC aggregates in compartmentalized microchambers. |
| 巻・号 | 124(2) |
| ページ | 234-241 |
| 公開日 | 2017-8-1 |
| DOI | 10.1016/j.jbiosc.2017.03.014 |
| PII | S1389-1723(16)30706-X |
| PMID | 28434976 |
| MeSH | Cell Count Cell Culture Techniques / instrumentation Cell Culture Techniques / methods* Cell Differentiation Fibronectins / metabolism Humans Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / metabolism Microfluidics / instrumentation Microfluidics / methods* |
| IF | 2.366 |
| 引用数 | 4 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | 201B7(HPS0063) |