Efficient Derivation and Banking of Clinical-grade Human Embryonic Stem Cell Lines in Accordance with Japanese Regulations

Overview

Journal Regen Ther

Date 2022 Nov 18

PMID 36397823

Authors

Kei Takada

Ryoko Nakatani

Emiko Moribe

Shizuka Yamazaki-Fujigaki

Mai Fujii

Masayo Furuta

Hirofumi Suemori

Eihachiro Kawase

Affiliations

Soon will be listed here.

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.

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