Human Fibrinogen for Maintenance and Differentiation of Induced Pluripotent Stem Cells in Two Dimensions and Three Dimensions

Jarel K. Gandhi, Travis Knudsen, Matthew Hill, Bhaskar Roy, Lori Bachman, Cynthia Pfannkoch-Andrews, Karina N. Schmidt, Muriel M. Metko, Michael John Ackerman, Zachary Resch, Jose S Pulido, Alan D Marmorstein

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Human fibrin hydrogels are a popular choice for use as a biomaterial within tissue engineered constructs because they are biocompatible, nonxenogenic, autologous use compatible, and biodegradable. We have recently demonstrated the ability to culture induced pluripotent stem cell (iPSC)-derived retinal pigment epithelium on fibrin hydrogels. However, iPSCs themselves have relatively few substrate options (e.g., laminin) for expansion in adherent cell culture for use in cell therapy. To address this, we investigated the potential of culturing iPSCs on fibrin hydrogels for three-dimensional applications and further examined the use of fibrinogen, the soluble precursor protein, as a coating substrate for traditional adherent cell culture. iPSCs successfully adhered to and proliferated on fibrin hydrogels. The two-dimensional culture with fibrinogen allows for immediate adaption of culture models to a nonxenogeneic model. Similarly, multiple commercially available iPSC lines adhered to and proliferated on fibrinogen coated surfaces. iPSCs cultured on fibrinogen expressed similar levels of the pluripotent stem cell markers SSea4 (98.7% ± 1.8%), Oct3/4 (97.3% ± 3.8%), TRA1-60 (92.2% ± 5.3%), and NANOG (96.0% ± 3.9%) compared with iPSCs on Geltrex. Using a trilineage differentiation assay, we found no difference in the ability of iPSCs grown on fibrinogen or Geltrex to differentiate to endoderm, mesoderm, or ectoderm. Finally, we demonstrated the ability to differentiate iPSCs to endothelial cells using only fibrinogen coated plates. On the basis of these data, we conclude that human fibrinogen provides a readily available and inexpensive alternative to laminin-based products for the growth, expansion, and differentiation of iPSCs for use in research and clinical cell therapy applications.

Original languageEnglish (US)
JournalStem Cells Translational Medicine
DOIs
StatePublished - Jan 1 2019

Fingerprint

Induced Pluripotent Stem Cells
Fibrinogen
Maintenance
Hydrogels
Fibrin
Laminin
Cell- and Tissue-Based Therapy
Cell Culture Techniques
Pluripotent Stem Cells
Endoderm
Ectoderm
Protein Precursors
Retinal Pigment Epithelium
Biocompatible Materials
Mesoderm
Endothelial Cells
Cell Line
Growth
Research

Keywords

  • Clinical translation
  • Endothelial cells
  • Induced pluripotent stem cells
  • Tissue engineering

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Human Fibrinogen for Maintenance and Differentiation of Induced Pluripotent Stem Cells in Two Dimensions and Three Dimensions. / Gandhi, Jarel K.; Knudsen, Travis; Hill, Matthew; Roy, Bhaskar; Bachman, Lori; Pfannkoch-Andrews, Cynthia; Schmidt, Karina N.; Metko, Muriel M.; Ackerman, Michael John; Resch, Zachary; Pulido, Jose S; Marmorstein, Alan D.

In: Stem Cells Translational Medicine, 01.01.2019.

Research output: Contribution to journalArticle

Gandhi, Jarel K. ; Knudsen, Travis ; Hill, Matthew ; Roy, Bhaskar ; Bachman, Lori ; Pfannkoch-Andrews, Cynthia ; Schmidt, Karina N. ; Metko, Muriel M. ; Ackerman, Michael John ; Resch, Zachary ; Pulido, Jose S ; Marmorstein, Alan D. / Human Fibrinogen for Maintenance and Differentiation of Induced Pluripotent Stem Cells in Two Dimensions and Three Dimensions. In: Stem Cells Translational Medicine. 2019.
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abstract = "Human fibrin hydrogels are a popular choice for use as a biomaterial within tissue engineered constructs because they are biocompatible, nonxenogenic, autologous use compatible, and biodegradable. We have recently demonstrated the ability to culture induced pluripotent stem cell (iPSC)-derived retinal pigment epithelium on fibrin hydrogels. However, iPSCs themselves have relatively few substrate options (e.g., laminin) for expansion in adherent cell culture for use in cell therapy. To address this, we investigated the potential of culturing iPSCs on fibrin hydrogels for three-dimensional applications and further examined the use of fibrinogen, the soluble precursor protein, as a coating substrate for traditional adherent cell culture. iPSCs successfully adhered to and proliferated on fibrin hydrogels. The two-dimensional culture with fibrinogen allows for immediate adaption of culture models to a nonxenogeneic model. Similarly, multiple commercially available iPSC lines adhered to and proliferated on fibrinogen coated surfaces. iPSCs cultured on fibrinogen expressed similar levels of the pluripotent stem cell markers SSea4 (98.7{\%} ± 1.8{\%}), Oct3/4 (97.3{\%} ± 3.8{\%}), TRA1-60 (92.2{\%} ± 5.3{\%}), and NANOG (96.0{\%} ± 3.9{\%}) compared with iPSCs on Geltrex. Using a trilineage differentiation assay, we found no difference in the ability of iPSCs grown on fibrinogen or Geltrex to differentiate to endoderm, mesoderm, or ectoderm. Finally, we demonstrated the ability to differentiate iPSCs to endothelial cells using only fibrinogen coated plates. On the basis of these data, we conclude that human fibrinogen provides a readily available and inexpensive alternative to laminin-based products for the growth, expansion, and differentiation of iPSCs for use in research and clinical cell therapy applications.",
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AU - Pfannkoch-Andrews, Cynthia

AU - Schmidt, Karina N.

AU - Metko, Muriel M.

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