Fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system.
Bioengineered functional cardiac tissue is expected to contribute to the repair of injured heart tissue. We previously developed cardiac cell sheets using mouse embryonic stem (mES) cell-derived cardiomyocytes, a system to generate an appropriate number of cardiomyocytes derived from ES cells and th...
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2012
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oai:doaj.org-article:551bb198e3024427abf1319d5f8118202021-11-18T08:04:17ZFabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system.1932-620310.1371/journal.pone.0052176https://doaj.org/article/551bb198e3024427abf1319d5f8118202012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23284924/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Bioengineered functional cardiac tissue is expected to contribute to the repair of injured heart tissue. We previously developed cardiac cell sheets using mouse embryonic stem (mES) cell-derived cardiomyocytes, a system to generate an appropriate number of cardiomyocytes derived from ES cells and the underlying mechanisms remain elusive. In the present study, we established a cultivation system with suitable conditions for expansion and cardiac differentiation of mES cells by embryoid body formation using a three-dimensional bioreactor. Daily conventional medium exchanges failed to prevent lactate accumulation and pH decreases in the medium, which led to insufficient cell expansion and cardiac differentiation. Conversely, a continuous perfusion system maintained the lactate concentration and pH stability as well as increased the cell number by up to 300-fold of the seeding cell number and promoted cardiac differentiation after 10 days of differentiation. After a further 8 days of cultivation together with a purification step, around 1 × 10(8) cardiomyocytes were collected in a 1-L bioreactor culture, and additional treatment with noggin and granulocyte colony stimulating factor increased the number of cardiomyocytes to around 5.5 × 10(8). Co-culture of mES cell-derived cardiomyocytes with an appropriate number of primary cultured fibroblasts on temperature-responsive culture dishes enabled the formation of cardiac cell sheets and created layered-dense cardiac tissue. These findings suggest that this bioreactor system with appropriate medium might be capable of preparing cardiomyocytes for cell sheet-based cardiac tissue.Katsuhisa MatsuuraMasanori WadaKanako KonishiMichi SatoUshio IwamotoYuko SatoAki TachibanaTetsutaro KikuchiTakahiro IwamiyaTatsuya ShimizuJun K YamashitaMasayuki YamatoNobuhisa HagiwaraTeruo OkanoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e52176 (2012) |
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Medicine R Science Q Katsuhisa Matsuura Masanori Wada Kanako Konishi Michi Sato Ushio Iwamoto Yuko Sato Aki Tachibana Tetsutaro Kikuchi Takahiro Iwamiya Tatsuya Shimizu Jun K Yamashita Masayuki Yamato Nobuhisa Hagiwara Teruo Okano Fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system. |
| description |
Bioengineered functional cardiac tissue is expected to contribute to the repair of injured heart tissue. We previously developed cardiac cell sheets using mouse embryonic stem (mES) cell-derived cardiomyocytes, a system to generate an appropriate number of cardiomyocytes derived from ES cells and the underlying mechanisms remain elusive. In the present study, we established a cultivation system with suitable conditions for expansion and cardiac differentiation of mES cells by embryoid body formation using a three-dimensional bioreactor. Daily conventional medium exchanges failed to prevent lactate accumulation and pH decreases in the medium, which led to insufficient cell expansion and cardiac differentiation. Conversely, a continuous perfusion system maintained the lactate concentration and pH stability as well as increased the cell number by up to 300-fold of the seeding cell number and promoted cardiac differentiation after 10 days of differentiation. After a further 8 days of cultivation together with a purification step, around 1 × 10(8) cardiomyocytes were collected in a 1-L bioreactor culture, and additional treatment with noggin and granulocyte colony stimulating factor increased the number of cardiomyocytes to around 5.5 × 10(8). Co-culture of mES cell-derived cardiomyocytes with an appropriate number of primary cultured fibroblasts on temperature-responsive culture dishes enabled the formation of cardiac cell sheets and created layered-dense cardiac tissue. These findings suggest that this bioreactor system with appropriate medium might be capable of preparing cardiomyocytes for cell sheet-based cardiac tissue. |
| format |
article |
| author |
Katsuhisa Matsuura Masanori Wada Kanako Konishi Michi Sato Ushio Iwamoto Yuko Sato Aki Tachibana Tetsutaro Kikuchi Takahiro Iwamiya Tatsuya Shimizu Jun K Yamashita Masayuki Yamato Nobuhisa Hagiwara Teruo Okano |
| author_facet |
Katsuhisa Matsuura Masanori Wada Kanako Konishi Michi Sato Ushio Iwamoto Yuko Sato Aki Tachibana Tetsutaro Kikuchi Takahiro Iwamiya Tatsuya Shimizu Jun K Yamashita Masayuki Yamato Nobuhisa Hagiwara Teruo Okano |
| author_sort |
Katsuhisa Matsuura |
| title |
Fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system. |
| title_short |
Fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system. |
| title_full |
Fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system. |
| title_fullStr |
Fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system. |
| title_full_unstemmed |
Fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system. |
| title_sort |
fabrication of mouse embryonic stem cell-derived layered cardiac cell sheets using a bioreactor culture system. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2012 |
| url |
https://doaj.org/article/551bb198e3024427abf1319d5f811820 |
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