Collagen vitrigels with low-fibril density enhance human embryonic stem cell-derived retinal pigment epithelial cell maturation

J Tissue Eng Regen Med. 2018 Mar;12(3):821-829. doi: 10.1002/term.2598. Epub 2017 Nov 17.


Structural and biochemical cues of extracellular matrix can substantially influence the differentiation and maturation of cultured retinal pigment epithelial (RPE) cells. In this study, thin collagen vitrigels were engineered to create collagen nanofibrillar structures of different fibril densities in an effort to evaluate the maturation of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells. The ultrastructure of the different collagen vitrigels was characterized by transmission electron microscopy, and the mechanical properties were evaluated by tensile testing. The pigmentation and polarization of cells, in addition to key RPE marker gene and protein expression levels, were analyzed to determine the differentiation of hESCs on the gels. The hESC-RPE differentiation was most significant in collagen vitrigels with low fibril density with mature collagen fibrils with diameter of around 60 nm and Young's modulus of 2.41 ± 0.13 MPa. This study provides insight into the influence of collagen nanofibrillar structures on hESC-RPE maturation and presents a potential bioengineered substratum for hESC-RPE for future preclinical and clinical applications.

Keywords: cell maturation; collagen vitrigel; epithelial cells; fibril density; retina; stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Cell Differentiation*
  • Cell Polarity / drug effects
  • Cell Shape / drug effects
  • Cells, Cultured
  • Collagen / pharmacology*
  • Cross-Linking Reagents / pharmacology
  • Epithelial Cells / cytology*
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Epithelial Cells / ultrastructure
  • Gels / pharmacology*
  • Gene Expression Regulation / drug effects
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / drug effects
  • Human Embryonic Stem Cells / metabolism
  • Human Embryonic Stem Cells / ultrastructure
  • Humans
  • Pigmentation / drug effects
  • Polyesters / pharmacology
  • Retinal Pigment Epithelium / cytology*
  • Vitrification*


  • Cross-Linking Reagents
  • Gels
  • Polyesters
  • polycaprolactone
  • Collagen