Dataset Information
Efficient derivation and banking of clinical-grade human embryonic stem cell lines in accordance with Japanese regulations.
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ABSTRACT: Introduction
We recently established clinical-grade human embryonic stem cell (hESC) line KthES11 in accordance with current good manufacturing practice standards in Japan. Despite this success, the establishment efficiency was very low at 7.1% in the first period.Methods
To establish clinical-grade hESC lines, we used xeno-free chemically defined medium StemFit AK03N with the LM-E8 fragments instead of feeder cells. The protocol was then optimized, especially in the early culture phase.Results
We established five hESC lines (KthES12, KthES13, KthES14, KthES15, and KthES16) with 45.5% efficiency. All five hESC lines showed typical hESC-like morphology, a normal karyotype, pluripotent state, and differentiation potential for all three germ layers. Furthermore, we developed efficient procedures to prepare master cell stocks for clinical-grade hESC lines and an efficient strategy for quality control testing.Conclusions
Our master cell stocks of hESC lines may contribute to therapeutic applications using human pluripotent stem cells in Japan and other countries.
SUBMITTER: Takada K
PROVIDER: S-EPMC9647332 | biostudies-literature | 2022 Dec
REPOSITORIES: biostudies-literature
Publications
Efficient derivation and banking of clinical-grade human embryonic stem cell lines in accordance with Japanese regulations.
Regenerative therapy 20221106
<h4>Introduction</h4>We recently established clinical-grade human embryonic stem cell (hESC) line KthES11 in accordance with current good manufacturing practice standards in Japan. Despite this success, the establishment efficiency was very low at 7.1% in the first period.<h4>Methods</h4>To establish clinical-grade hESC lines, we used xeno-free chemically defined medium StemFit AK03N with the LM-E8 fragments instead of feeder cells. The protocol was then optimized, especially in the early cultur ...[more]