Reference - Detail
| RRC ID | 45019 |
|---|---|
| Author | Ohnuma K, Fujiki A, Yanagihara K, Tachikawa S, Hayashi Y, Ito Y, Onuma Y, Chan T, Michiue T, Furue MK, Asashima M. |
| Title | Enzyme-free passage of human pluripotent stem cells by controlling divalent cations. |
| Journal | Sci Rep |
| Abstract |
Enzymes used for passaging human pluripotent stem cells (hPSCs) digest cell surface proteins, resulting in cell damage. Moreover, cell dissociation using divalent cation-free solutions causes apoptosis. Here we report that Mg(2+) and Ca(2+) control cell-fibronectin and cell-cell binding of hPSCs, respectively, under feeder- and serum-free culture conditions without enzyme. The hPSCs were detached from fibronectin-, vitronectin- or laminin-coated dishes in low concentrations of Mg(2+) and remained as large colonies in high concentrations of Ca(2+). Using enzyme-free solutions containing Ca(2+) without Mg(2+), we successfully passaged hPSCs as large cell clumps that showed less damage than cells passaged using a divalent cation-free solution or dispase. Under the same conditions, the undifferentiated and early-differentiated cells could also be harvested as a cell sheet without being split off. Our enzyme-free passage of hPSCs under a serum- and feeder-free culture condition reduces cell damage and facilitates easier and safer cultures of hPSCs. |
| Volume | 4 |
| Pages | 4646 |
| Published | 2014-4-11 |
| DOI | 10.1038/srep04646 |
| PII | srep04646 |
| PMID | 24721898 |
| PMC | PMC3983606 |
| MeSH | Calcium / chemistry Calcium / metabolism Cations, Divalent / chemistry Cations, Divalent / metabolism* Cell Differentiation Cell Line Cell Proliferation Endopeptidases / metabolism Fibronectins / metabolism Humans Karyotyping Magnesium / chemistry Magnesium / metabolism Pluripotent Stem Cells / cytology* Pluripotent Stem Cells / metabolism |
| IF | 3.998 |
| Times Cited | 18 |
| WOS Category | MULTIDISCIPLINARY SCIENCES |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 8 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | 201B7(HPS0063) 253G1(HPS0002) |