Methods in Molecular Biology (2022) 2436: 113–125
DOI 10.1007/7651_2021_425
© Springer Science+Business Media, LLC 2021
Published online: 15 September 2021
Large-Scale Expansion of Umbilical Cord Mesenchymal
Stem Cells with Microcarrier Tablets in Bioreactor
Huanye Xu, Zhongxiao Cong, Yuanyuan Zhang, Wei Liu, Xiaojun Yan,
and Yanan Du
Abstract
Mesenchymal stem cells show great potential in tissue engineering and cell-based therapies. This protocol
demonstrates the use of 3D TableTrix® microcarrier tablets for large-scale manufacturing of human
umbilical cord mesenchymal stem cells (hUCMSCs) in a 5-L stirred tank bioreactor. 3D TableTrix®
microcarrier tablets readily disperse into tens of thousands of porous microcarriers to simplify cell seeding,
and 3D FloTrix® vivaSPIN bioreactor could automate, monitor, and control the entire culture process. 3D
TableTrix® microcarriers could also be fully dissolved upon adding dissolution reagent to gently harvest the
expanded cells at a high recovery rate. With this protocol, more than 109 cells could be produced in a 5-L
bioreactor.
Key words 3D cell culture, Bioreactor, Dissolvable microcarrier, Large-scale expansion, Mesenchymal
stem cells
1
Introduction
Umbilical cord mesenchymal stem cells (UCMSCs) have high
capacity of multilineage differentiation and immunomodulatory
effects, which show promising prospects in hematopoietic function
recovery, tissue injury repair and regeneration [1]. UCMSCs have
been greatly applied in tissue engineering and cell-based therapies
for stroke, spinal cord injury, neurodegenerative diseases, etc.
[2]. Human UCMSCs (hUCMSCs), which could be easily isolated
from umbilical cord, with minimal invasiveness to both neonate or
mother, get more attention due to their high multiplication capac-
ity, low immunogenicity, as well as little ethical concerns [3]. Now-
adays, the demand for UCMSCs is exceeding supply because the
required dose for clinical efficacy is high, usually ranging from 40 to
100 million cells [4]. Traditional in vitro monolayer cell culture
methods under static conditions are labor intensive and have lim-
ited production capacity and efficiency [5]. To produce huge
Huanye Xu and Zhongxiao Cong contributed equally to this work.
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