Open AccessA fully defined static suspension culture system for large-scale human embryonic stem cell production
Author(s) -
Xia Li,
Ruisong Ma,
Qi Gu,
Liping Liang,
Lei Wang,
Ying Zhang,
Xianning Wang,
Xin Liu,
Zhongwen Li,
Jie Fang,
Jun Wu,
Yukai Wang,
Wei Li,
Baoyang Hu,
Liu Wang,
Qi Zhou,
Jie Hao
Publication year - 2018
Publication title -
cell death and disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.482
H-Index - 111
ISSN - 2041-4889
DOI - 10.1038/s41419-018-0863-8
Subject(s) - embryonic stem cell , cell culture , regenerative medicine , suspension culture , microbiology and biotechnology , stem cell , biology , computer science , biomedical engineering , medicine , biochemistry , genetics , gene
Human embryonic stem cells (hESCs) play an important role in regenerative medicine due to their potential to differentiate into various functional cells. However, the conventional adherent culture system poses challenges to mass production of high-quality hESCs. Though scientists have made many attempts to establish a robust and economical hESC suspension culture system, there are existing limitations, including suboptimal passage methods and shear force caused by dynamic stirring. Here, we report on an efficient large-scale culture system, which enables long-term, GMP grade, single-cell inoculation, and serial expansion of hESCs with a yield of about 1.5 × 10 9 cells per 1.5-L culture, while maintaining good pluripotency. The suspension culture system was enlarged gradually from a 100-mm dish to a 1.8-L culture bag with methylcellulose involvement to avoid sphere fusion. Under the optimal experimental protocol, this 3D system resolves current problems that limit mass production and clinical application of hESCs, and thus can be used in commercial-level hESC production for cell therapy and pharmaceutics screening in the future.