Review
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jan 26, 2020; 12(1): 25-34
Published online Jan 26, 2020. doi: 10.4252/wjsc.v12.i1.25
Inducing human induced pluripotent stem cell differentiation through embryoid bodies: A practical and stable approach
Ning-Ning Guo, Li-Ping Liu, Yun-Wen Zheng, Yu-Mei Li
Ning-Ning Guo, Li-Ping Liu, Yun-Wen Zheng, Yu-Mei Li, Institute of Regenerative Medicine, Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang 212001, Jiangsu Province, China
Yun-Wen Zheng, Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery, University of Tsukuba Faculty of Medicine, Tsukuba, Ibaraki 305-8575, Japan
Yun-Wen Zheng, Yokohama City University School of Medicine, Yokohama, Kanagawa 234-0006, Japan
Yun-Wen Zheng, Division of Regenerative Medicine, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, the University of Tokyo, Tokyo 108-8639, Japan
Author contributions: Guo NN conceived the study and drafted the manuscript; Liu LP, Zheng YW and Li YM contributed in discussions on the subject and reviewing the final manuscript text; all the authors approved the final version of the article; Guo NN and Liu LP contributed equally to this work; Li YM and Zheng YW are senior authors and co-correspondent to this work.
Supported by National Natural Science Foundation of China, No. 81770621, No. 81573053; Ministry of Education, Culture, Sports, Science, and Technology of Japan, KAKENHI, No. 16K15604, No. 18H02866; Natural Science Foundation of Jiangsu Province, No. BK20180281.
Conflict-of-interest statement: No potential conflict of interest.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Yun-Wen Zheng, PhD, Associate Professor, Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery, University of Tsukuba Faculty of Medicine,Tennodai 1-1-1, Tsukuba, Ibaraki 305-8575, Japan. ywzheng@md.tsukuba.ac.jp
Received: April 1, 2019
Peer-review started: April 3, 2019
First decision: August 23,2019
Revised: September 30, 2019
Accepted: December 13, 2019
Article in press: December 13, 2019
Published online: January 26, 2020
Abstract

Human induced pluripotent stem cells (hiPSCs) are invaluable resources for producing high-quality differentiated cells in unlimited quantities for both basic research and clinical use. They are particularly useful for studying human disease mechanisms in vitro by making it possible to circumvent the ethical issues of human embryonic stem cell research. However, significant limitations exist when using conventional flat culturing methods especially concerning cell expansion, differentiation efficiency, stability maintenance and multicellular 3D structure establishment, differentiation prediction. Embryoid bodies (EBs), the multicellular aggregates spontaneously generated from iPSCs in the suspension system, might help to address these issues. Due to the unique microenvironment and cell communication in EB structure that a 2D culture system cannot achieve, EBs have been widely applied in hiPSC-derived differentiation and show significant advantages especially in scaling up culturing, differentiation efficiency enhancement, ex vivo simulation, and organoid establishment. EBs can potentially also be used in early prediction of iPSC differentiation capability. To improve the stability and feasibility of EB-mediated differentiation and generate high quality EBs, critical factors including iPSC pluripotency maintenance, generation of uniform morphology using micro-pattern 3D culture systems, proper cellular density inoculation, and EB size control are discussed on the basis of both published data and our own laboratory experiences. Collectively, the production of a large quantity of homogeneous EBs with high quality is important for the stability and feasibility of many PSCs related studies.

Keywords: Induced pluripotent stem cells, Suspension culture, Embryoid body, Early prediction, Committed differentiation, Heterogeneity, Three-dimensional culture, Scaling-up, Quality control

Core tip:Embryoid body (EB) mediated induced pluripotent stem cell (iPSC) differentiation shows great advantages in culture scale-up, differentiation efficiency improvement, ex vivo simulation and organoid establishment. To improve the stability and feasibility of high quality EB generation, factors including iPSC pluripotency maintenance, generation of uniform morphology using micro-pattern 3D culture systems, proper cellular density inoculation and EB size control need to be considered.