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

• Published in: Stem cells translational medicine Vol. 8; no. 6; pp. 512 - 521
• Main Authors: Gandhi, Jarel K., Knudsen, Travis, Hill, Matthew, Roy, Bhaskar, Bachman, Lori, Pfannkoch‐Andrews, Cynthia, Schmidt, Karina N., Metko, Muriel M., Ackerman, Michael J., Resch, Zachary, Pulido, Jose S., Marmorstein, Alan D.
• Format: Journal Article
• Language: English
• Published: United States Oxford University Press 01.06.2019; John Wiley and Sons Inc
• Subjects: Antigens, CD - metabolism; Autografts; Biodegradability; Biomaterials; Cadherins - metabolism; Cell culture; Cell Culture Techniques - methods; Cell differentiation; Cell Differentiation - drug effects; Clinical translation; Cloning; Coatings; Data analysis; Ectoderm; Endoderm; Endothelial cells; Endothelial Cells - cytology; Endothelial Cells - metabolism; Epithelium; Fibrin; Fibrin - chemistry; Fibrin - pharmacology; Fibrinogen; Fibrinogen - analysis; Fibrinogen - isolation & purification; Humans; Hydrogels; Hydrogels - chemistry; Induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Laboratories; Laminin; Mesoderm; Nanog Homeobox Protein - metabolism; Oct-4 protein; Octamer Transcription Factor-3 - metabolism; Pluripotency; Pluripotent Stem Cells; Proteins; Retina; Retinal pigment epithelium; Scholarships & fellowships; Stem cells; Tissue engineering; Clinical translation; Induced pluripotent stem cells; Tissue engineering; Endothelial cells
• ISSN: 2157-6564; 2157-6580
• DOI: 10.1002/sctm.18-0189

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. Stem Cells Translational Medicine 2019;8:512–521 Induced pluripotent stem cells cultured on fibrinogen coated plates successfully maintain pluripotency, differentiate to the three germlayers, and can be specifically differentiated to endothelial cells.