Affinity Selection of FGF2-Binding Heparan Sulfates for Ex Vivo Expansion of Human Mesenchymal Stem Cells

Sampath Jeewantha Wijesinghe, Ling Ling, Sadasivam Murali, Yeong Hui Qing, Simon F R Hinkley, Susan M. Carnachan, Tracey J. Bell, Kunchithapadam Swaminathan, James H. Hui, Andre J van Wijnen, Victor Nurcombe, Simon M. Cool

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
JournalJournal of Cellular Physiology
DOIs
StateAccepted/In press - 2016

Fingerprint

Heparitin Sulfate
Fibroblast Growth Factor 2
Stem cells
Mesenchymal Stromal Cells
Fibroblast Growth Factor 1
Cell culture
Ligands
Stem Cell Niche
Fibroblast Growth Factor Receptors
Surface Antigens
Cell- and Tissue-Based Therapy
Cell proliferation
Chromatography
Bioactivity
Heparin
Intercellular Signaling Peptides and Proteins
Swine
Cell Count
Hot Temperature
Scalability

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Wijesinghe, S. J., Ling, L., Murali, S., Qing, Y. H., Hinkley, S. F. R., Carnachan, S. M., ... Cool, S. M. (Accepted/In press). Affinity Selection of FGF2-Binding Heparan Sulfates for Ex Vivo Expansion of Human Mesenchymal Stem Cells. Journal of Cellular Physiology. https://doi.org/10.1002/jcp.25454

Affinity Selection of FGF2-Binding Heparan Sulfates for Ex Vivo Expansion of Human Mesenchymal Stem Cells. / Wijesinghe, Sampath Jeewantha; Ling, Ling; Murali, Sadasivam; Qing, Yeong Hui; Hinkley, Simon F R; Carnachan, Susan M.; Bell, Tracey J.; Swaminathan, Kunchithapadam; Hui, James H.; van Wijnen, Andre J; Nurcombe, Victor; Cool, Simon M.

In: Journal of Cellular Physiology, 2016.

Research output: Contribution to journalArticle

Wijesinghe, SJ, Ling, L, Murali, S, Qing, YH, Hinkley, SFR, Carnachan, SM, Bell, TJ, Swaminathan, K, Hui, JH, van Wijnen, AJ, Nurcombe, V & Cool, SM 2016, 'Affinity Selection of FGF2-Binding Heparan Sulfates for Ex Vivo Expansion of Human Mesenchymal Stem Cells', Journal of Cellular Physiology. https://doi.org/10.1002/jcp.25454
Wijesinghe, Sampath Jeewantha ; Ling, Ling ; Murali, Sadasivam ; Qing, Yeong Hui ; Hinkley, Simon F R ; Carnachan, Susan M. ; Bell, Tracey J. ; Swaminathan, Kunchithapadam ; Hui, James H. ; van Wijnen, Andre J ; Nurcombe, Victor ; Cool, Simon M. / Affinity Selection of FGF2-Binding Heparan Sulfates for Ex Vivo Expansion of Human Mesenchymal Stem Cells. In: Journal of Cellular Physiology. 2016.
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