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Efficient Large-Scale 2D Culture System for Human Induced Pluripotent Stem Cells and Differentiated Cardiomyocytes
- Source :
- Stem Cell Reports, Stem Cell Reports, Vol 9, Iss 5, Pp 1406-1414 (2017)
- Publication Year :
- 2017
- Publisher :
- Elsevier, 2017.
-
Abstract
- Summary Cardiac regenerative therapies utilizing human induced pluripotent stem cells (hiPSCs) are hampered by ineffective large-scale culture. hiPSCs were cultured in multilayer culture plates (CPs) with active gas ventilation (AGV), resulting in stable proliferation and pluripotency. Seeding of 1 × 106 hiPSCs per layer yielded 7.2 × 108 hiPSCs in 4-layer CPs and 1.7 × 109 hiPSCs in 10-layer CPs with pluripotency. hiPSCs were sequentially differentiated into cardiomyocytes (CMs) in a two-dimensional (2D) differentiation protocol. The efficiency of cardiac differentiation using 10-layer CPs with AGV was 66%–87%. Approximately 6.2–7.0 × 108 cells (4-layer) and 1.5–2.8 × 109 cells (10-layer) were obtained with AGV. After metabolic purification with glucose- and glutamine-depleted and lactate-supplemented media, a massive amount of purified CMs was prepared. Here, we present a scalable 2D culture system using multilayer CPs with AGV for hiPSC-derived CMs, which will facilitate clinical applications for severe heart failure in the near future.<br />Graphical Abstract<br />Highlights • Efficient mass production of hiPSCs by multilayer culture plates with AGV • Efficient mass production of hiPSC-CMs using a massive 2D culture system with AGV • Mass production of pure hiPSC-CMs via metabolic selection<br />In this article, Fujita and colleagues show that human induced pluripotent stem cells (hiPSCs) and hiPSC-derived cardiomyocytes (hiPSC-CMs) are stably and efficiently produced using multilayer culture plates with an active gas ventilation system because of stable CO2 and O2 concentration in culture media. Large numbers of obtained hiPSC-CMs are metabolically purified under glucose- and glutamine-depleted and lactate-supplemented conditions.
- Subjects :
- 0301 basic medicine
Culture plates
purification
Cardiac differentiation
Induced Pluripotent Stem Cells
Primary Cell Culture
iPSCs
regenerative medicine
cardiomyocytes
Biology
Biochemistry
03 medical and health sciences
Report
2D culture
Genetics
Humans
Myocytes, Cardiac
Human Induced Pluripotent Stem Cells
lcsh:QH301-705.5
Cells, Cultured
Cell Proliferation
lcsh:R5-920
Cell Differentiation
Cell Biology
Cell biology
Culture Media
030104 developmental biology
lcsh:Biology (General)
large scale
lcsh:Medicine (General)
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 22136711
- Volume :
- 9
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Stem Cell Reports
- Accession number :
- edsair.doi.dedup.....9c7a18783a66591db6a05c8dbc480bd8