Efficient derivation and banking of clinical-grade human embryonic stem cell lines in accordance with Japanese regulations.

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 dev

SUBMITTER: Takada K 

PROVIDER: S-EPMC9647332 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature