Maintenance of human pluripotent stem cells using 4SP-hFGF2-secreting STO cells

• Published in: Stem cell research 2011-11, Vol.7 (3), p.210-218
• Main Authors: Lee, Won-Young, Kim, Jumi, Gil, Chang-Hyun, Lee, Jae-Ho, Song, Hyuk, Kim, Jae-Hwan, Chung, Hyung-Min
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Blocking - pharmacology; Base Sequence; Cell Culture Techniques - methods; Cell Differentiation - drug effects; Cell Line; Embryonic Stem Cells - cytology; Embryonic Stem Cells - drug effects; Embryonic Stem Cells - metabolism; Fibroblast Growth Factor 2 - immunology; Fibroblast Growth Factor 2 - secretion; Fibroblast Growth Factor 4 - metabolism; Genetic Vectors - genetics; Germ Layers - cytology; Germ Layers - drug effects; Humans; Immunoglobulin G - pharmacology; Mice; Molecular Sequence Data; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - drug effects; Pluripotent Stem Cells - metabolism; Protein Sorting Signals; Recombinant Fusion Proteins - secretion; Subcellular Fractions - drug effects; Subcellular Fractions - metabolism
• ISSN: 1873-5061; 1876-7753
• DOI: 10.1016/j.scr.2011.05.004

Human embryonic stem cells (hESCs) are typically cultured on fibroblast feeder cells or in fibroblast conditioned medium supplemented with fibroblast growth factor 2 (FGF2, also known as bFGF). FGF signaling appears to be important for hESC self-renewal and is required to enable the culture of hESCs in an undifferentiated state. In this study, we generated a transgenic fibroblast feeder line stably expressing a secretable FGF4 signal peptide tagged hFGF2 (4SP-hFGF2). The expression of this transgene functionally replaced the requirement for exogenous FGF2 when using these cells as feeders for the maintenance of hESCs. Under these conditions, hESCs maintained the typical marker of pluripotency assessed after long term culture, while still retaining the capacity for differentiation to all three germ layers. This transgene could be applied to mass produce 4SP-hFGF2 protein, serving to be an economical and effective strategy for culturing pluripotent stem cells as feeder cells. ► FGF signaling appears to be important in hESC self-renewal. ► FGF2 lacks a signal sequence as it's released from damaged or dead cells. ► We made FGF2 secreted cells, using a lentiviral system tagged FGF4 signal peptide. ► Two types of hESC lines proliferated without differentiation on 4SP-hFGF2 STO cells. ► 4SP-hFGF2 cells may be a simple and economic feeder system for hESCs.