A New Feeder-Free Technique to Expand Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

Mark Denham 1, Jessie Leung 1, Cheryl Tay 1, 2, Raymond C.B. Wong 3, Peter Donovan 4, Mirella Dottori 1, 5, Alice Pébay 1, 5, *
Centre for Neuroscience, The University of Melbourne, Parkville, VIC 3010, Australia
Monash Institute of Medical Research, Monash University, Clayton, VIC 3168, Australia
Department of Biological Chemistry
Department of Development and Cell Biology University of California Irvine, Irvine, CA, 92697, USA
Department of Pharmacology, The University of Melbourne, Parkville, VIC 3010, Australia

© Pébay et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Centre for Neuroscience, The University of Melbourne, Parkville, VIC 3010, Australia.


The optimal maintenance of human embryonic stem cells (hESC) is generally observed in the presence of a feeder-layer of mouse embryonic fibroblasts in a serum-containing medium. Various approaches are now available to remove the feeder requirement. Today, the best feeder-free system for the maintenance of hESC and induced pluripotent stem (iPS) cells is based on a serum-replacement medium on a matrice of Matrigel. Some reports have also shown feederfree maintenance of hESC in the presence of laminin, fibronectin, vitronectin or collagen IV. However these combinations are expensive. Here we describe an alternative to the current feeder-free short-term amplification of hESC and iPS cells using culture grade plastic dishes pre-coated with 10-20% fetal calf serum. hESC retain expression of various stem cell markers and also retain the ability to differentiate . Similar results were observed with human iPS cells. This feeder-free culture system is reliable, convenient and allows for the rapid amplification of hESC and iPS cells to numbers suitable for many cell biology techniques.