Affinity Selection of FGF2-Binding Heparan Sulfates for Ex Vivo Expansion of Human Mesenchymal Stem Cells.
The future of human mesenchymal stem cells (hMSCs) as a successful cell therapy relies on bioprocessing strategies to improve the scalability of these cells without compromising their therapeutic ability. The culture-expansion of hMSCs can be enhanced by supplementation with growth factors, particularly fibroblast growth factor 2 (FGF2). The biological activity of FGF2 is controlled through interactions with heparan sulfate (HS) that facilitates ligand-receptor complex formation. We previously reported on an FGF2-interacting HS variant (termed HS2) isolated from embryonic tissue by anionic exchange chromatography that increased the proliferation and potency of hMSCs. Here, we detail the isolation of an FGF2 affinity-purified HS variant (HS8) using a scalable platform technology previously employed to generate HS variants with increased affinity for BMP-2 or VEGF165. This process used a peptide sequence derived from the heparin-binding domain of FGF2 as a substrate to affinity-isolate HS8 from a commercially available source of porcine mucosal HS. Our data show that HS8 binds to FGF2 with higher affinity than to FGF1, FGF7, BMP2, PDGF-BB, or VEGF165. Also, HS8 protects FGF2 from thermal destabilization and increases FGF signaling and hMSC proliferation through FGF receptor 1. Long-term supplementation of cultures with HS8 increased both hMSC numbers and their colony-forming efficiency without adversely affecting the expression of hMSC-related cell surface antigens. This strategy further exemplifies the utility of affinity-purifying HS variants against particular ligands important to the stem cell microenvironment and advocates for their addition as adjuvants for the culture-expansion of hMSCs destined for cellular therapy. J. Cell. Physiol. 232: 566-575, 2017. © 2016 Wiley Periodicals, Inc.
Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore.
Sampath Jeewantha
SJ
Wijesinghe
Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore.
Ling
L
Ling
Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore.
Sadasivam
S
Murali
Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore.
Yeong Hui
YH
Qing
The Ferrier Research Institute, Victoria University of Wellington, Lower Hutt, New Zealand.
Simon F R
SF
Hinkley
The Ferrier Research Institute, Victoria University of Wellington, Lower Hutt, New Zealand.
Susan M
SM
Carnachan
The Ferrier Research Institute, Victoria University of Wellington, Lower Hutt, New Zealand.
Tracey J
TJ
Bell
Department of Biological Sciences, National University of Singapore, Singapore.
Kunchithapadam
K
Swaminathan
Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
James H
JH
Hui
Department of Orthopedic Surgery and Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota.
Andre J
AJ
van Wijnen
Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore.
Victor
V
Nurcombe
Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore.|Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Simon M
SM
Cool
D000595:Amino Acid Sequence; D000925:Anticoagulants / Q000494:pharmacology; D049109:Cell Proliferation; D002846:Chromatography, Affinity; D004187:Disaccharides / Q000032:analysis; D015951:Factor Xa / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000378:metabolism*; D006497:Heparan Sulfate / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D055550:Protein Stability / Q000187:drug effects; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
D000925:Anticoagulants; D004187:Disaccharides; D015951:Factor Xa; D016222:Fibroblast Growth Factor 2; D006497:Heparan Sulfate; D010455:Peptides; D051496:Receptor, Fibroblast Growth Factor, Type 1