| RRC ID |
45468
|
| 著者 |
Takenaka C, Miyajima H, Yoda Y, Imazato H, Yamamoto T, Gomi S, Ohshima Y, Kagawa K, Sasaki T, Kawamata S.
|
| タイトル |
Controlled Growth and the Maintenance of Human Pluripotent Stem Cells by Cultivation with Defined Medium on Extracellular Matrix-Coated Micropatterned Dishes.
|
| ジャーナル |
PLoS One
|
| Abstract |
Here, we introduce a new serum-free defined medium (SPM) that supports the cultivation of human pluripotent stem cells (hPSCs) on recombinant human vitronectin-N (rhVNT-N)-coated dishes after seeding with either cell clumps or single cells. With this system, there was no need for an intervening sequential adaptation process after moving hPSCs from feeder layer-dependent conditions. We also introduce a micropatterned dish that was coated with extracellular matrix by photolithographic technology. This procedure allowed the cultivation of hPSCs on 199 individual rhVNT-N-coated small round spots (1 mm in diameter) on each 35-mm polystyrene dish (termed "patterned culture"), permitting the simultaneous formation of 199 uniform high-density small-sized colonies. This culture system supported controlled cell growth and maintenance of undifferentiated hPSCs better than dishes in which the entire surface was coated with rhVNT-N (termed "non-patterned cultures"). Non-patterned cultures produced variable, unrestricted cell proliferation with non-uniform cell growth and uneven densities in which we observed downregulated expression of some self-renewal-related markers. Comparative flow cytometric studies of the expression of pluripotency-related molecules SSEA-3 and TRA-1-60 in hPSCs from non-patterned cultures and patterned cultures supported this concept. Patterned cultures of hPSCs allowed sequential visual inspection of every hPSC colony, giving an address and number in patterned culture dishes. Several spots could be sampled for quality control tests of production batches, thereby permitting the monitoring of hPSCs in a single culture dish. Our new patterned culture system utilizing photolithography provides a robust, reproducible and controllable cell culture system and demonstrates technological advantages for the mass production of hPSCs with process quality control.
|
| 巻・号 |
10(6)
|
| ページ |
e0129855
|
| 公開日 |
2015-1-1
|
| DOI |
10.1371/journal.pone.0129855
|
| PII |
PONE-D-15-10620
|
| PMID |
26115194
|
| PMC |
PMC4483150
|
| MeSH |
Animals
Cell Culture Techniques* / instrumentation
Cell Culture Techniques* / methods
Cell Differentiation
Cell Line
Cell Proliferation
Cell Self Renewal
Cryopreservation
Culture Media
Embryonic Stem Cells / cytology
Embryonic Stem Cells / metabolism
Extracellular Matrix / metabolism
Feeder Cells
Genomic Instability
Humans
Induced Pluripotent Stem Cells / cytology
Induced Pluripotent Stem Cells / metabolism
Karyotyping
Pluripotent Stem Cells / cytology*
Pluripotent Stem Cells / metabolism
|
| IF |
2.74
|
| 引用数 |
9
|
|
WOS 分野
|
MULTIDISCIPLINARY SCIENCES
|
| リソース情報 |
| ヒト・動物細胞 |
KhES-1(HES0001)
201B7(HPS0063) |
