Laminin as a Potent Substrate for Large-Scale Expansion of Human Induced Pluripotent Stem Cells in a Closed Cell Expansion System

Stem Cells International
Fernanda C Paccola MesquitaDoris A Taylor

Abstract

The number of high-quality cells required for engineering an adult human-sized bioartificial organ is greater than one billion. Until the emergence of induced pluripotent stem cells (iPSCs), autologous cell sources of this magnitude and with the required complexity were not available. Growing this number of cells in a traditional 2D cell culture system requires extensive time, resources, and effort and does not always meet clinical requirements. The use of a closed cell culture system is an efficient and clinically applicable method that can be used to expand cells under controlled conditions. We aimed to use the Quantum Cell Expansion System (QES) as an iPSC monolayer-based expansion system. Human iPSCs were expanded (up to 14-fold) using the QES on two different coatings (laminin 521 (LN521) and vitronectin (VN)), and a karyotype analysis was performed. The cells were characterized for spontaneous differentiation and pluripotency by RT-PCR and flow cytometry. Our results demonstrated that the QES provides the necessary environment for exponential iPSC growth, reaching 689.75 × 106 ± 86.88 × 106 in less than 7 days using the LN521 coating with a population doubling level of 3.80 ± 0.19. The same result was not observed when VN...Continue Reading

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Citations

Aug 4, 2020·Journal of Tissue Engineering·Camila Hochman-MendezAntonio Carlos Campos de Carvalho
Oct 23, 2019·Frontiers in Bioengineering and Biotechnology·Fei Tan, Mohamed Al-Rubeai
Nov 2, 2019·The Journal of Thoracic and Cardiovascular Surgery·Doris A TaylorLuiz C Sampaio
Aug 14, 2021·Cardiovascular Research·Doris A TaylorUNKNOWN Cardiovascular Cell Therapy Research Network (CCTRN)
Nov 3, 2021·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Araceli Rivera-OrdazLorenza Lazzari

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Methods Mentioned

BETA
PCR
electrophoresis
flow cytometry

Software Mentioned

GraphPad
FlowJo
GraphPad Prism®

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