| RRC ID |
45019
|
| 著者 |
Ohnuma K, Fujiki A, Yanagihara K, Tachikawa S, Hayashi Y, Ito Y, Onuma Y, Chan T, Michiue T, Furue MK, Asashima M.
|
| タイトル |
Enzyme-free passage of human pluripotent stem cells by controlling divalent cations.
|
| ジャーナル |
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.
|
| 巻・号 |
4
|
| ページ |
4646
|
| 公開日 |
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
|
| 引用数 |
18
|
|
WOS 分野
|
MULTIDISCIPLINARY SCIENCES
|
| リソース情報 |
| ヒト・動物細胞 |
201B7(HPS0063)
253G1(HPS0002) |
