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

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. stem cells translational medicine 2019;8:512–521

Increasing salinity of fibrinogen solvent generates stable fibrin hydrogels for cell delivery or tissue engineering

“…Recently, several studies have sought to identify materials capable of supporting iPSC for expansion and tissue engineering. Fibrin(ogen) has been found to be capable of this maintenance [ 34 , 35 ]. Because our group is interested in 3D stem cell differentiations with multiple stem cell types, we sought to corroborate these findings and determine if gel structure influences 3D iPSC culture.…”

Cyclically stretched ACL fibroblasts emigrating from spheroids adapt their cytoskeleton and ligament-related expression profile

“…Hence, chambers were coated with Geltrex® to guarantee a quick adherence of the spheroids since the pure silicone does not provide sufficient binding motifs. Geltrex® did not affect the differentiation capability of pluripotent stem cells in other studies (Gandhi et al 2019 ). Not only a quick adherence is guaranteed but also the maintenance of the ligament-specific characteristics, such as expression of collagen type I, DCN, TNC, MKX, and TNMD.…”

Thalidomide Inhibits Human iPSC Mesendoderm Differentiation by Modulating CRBN-dependent Degradation of SALL4

“…Thus we employed a 3D culture system amenable to mesoderm differentiation. Fibrin encapsulation supports maintenance of hPSCs 66 , cardiac mesoderm differentiation of hPSCs 67 , and LPM differentiation and subsequent limb-regenerating capability of mESCs 26 . Fibrin encapsulation here maintained the pluripotency of hiPSCs (Supplementary Table 2) and supported LPM differentiation with similar magnitude and time-course of marker expression as 2D culture (Fig.…”