Int. J. Dev. Biol. 52: 353 - 363 (2008)

https://doi.org/10.1387/ijdb.082590le

Vol 52, Issue 4

Comparative study of mouse and human feeder cells for human embryonic stem cells

Original Article | Published: 1 April 2008

Livia Eiselleova^1,4, Iveta Peterkova⁴, Jakub Neradil¹, Iva Slaninova¹, Ales Hamp^l,1,2,3 and Petr Dvorak*^,1,2,3

¹Department of Biology, Faculty of Medicine, Masaryk University, Brno, ²Department of Molecular Embryology, Institute of Experimental Medicine AS CR, Brno, ³Center for Cell Therapy and Tissue Repair, Charles University, Prague and ⁴Center for Chemical Genetics, Masaryk University, Brno, Czech Republic

Abstract

Various types of feeder cells have been adopted for the culture of human embryonic stem cells (hESCs) to improve their attachment and provide them with stemness-supporting factors. However, feeder cells differ in their capacity to support the growth of undifferentiated hESCs. Here, we compared the expression and secretion of four well-established regulators of hESC pluripotency and/or differentiation among five lines of human foreskin fibroblasts and primary mouse embryonic fibroblasts throughout a standard hESC culture procedure. We found that human and mouse feeder cells secreted comparable levels of TGFbeta1. However, mouse feeder cells secreted larger quantities of activin A than human feeder cells. Conversely, FGF-2, which was produced by human feeder cells, could not be detected in culture media from mouse feeder cells. The quantity of BMP-4 was at about the level of detectability in media from all feeder cell types, although BMP-4 dimers were present in all feeder cells. Production of TGFbeta1, activin A, and FGF-2 varied considerably among the human-derived feeder cell lines. Low- and high-producing human feeder cells as well as mouse feeder cells were evaluated for their ability to support the undifferentiated growth of hESCs. We found that a significantly lower proportion of hESCs maintained on human feeder cell types expressed SSEA3, an undifferentiated cell marker. Moreover, SSEA3 expression and thus the pluripotent hESC compartment could be partially rescued by addition of activin A. Cumulatively, these results suggest that the ability of a feeder layer to promote the undifferentiated growth of hESCs is attributable to its characteristic growth factor production.

Keywords

human embryonic stem cell, growth factor production, undifferentiated growth