Sphere formation from corneal keratocytes and phenotype specific markers

Sherri Gae Scott, Albert Jun, Shukti Chakravarti

Research output: Contribution to journalArticle

Abstract

The keratocytes are specialized mesenchymal cells that produce and maintain the extracellular matrix of the corneal stroma. With a typical dendritic and flattened appearance, these cells can morph into fibroblasts and myofibroblasts upon injury, and produce abnormal or fibrotic extracellular matrices detrimental to corneal transparency. Insights into mechanisms that regulate these phenotypic switches and optimal culture conditions that preserve the keratocyte phenotype are important for tissue engineering of the corneal stroma. Like other cell types with self-renewing capacity, keratocytes can form spheres in culture. Here we investigated human and bovine keratocytes with respect to their sphere forming capabilities, and sought to identify potentially distinguishing markers for the keratocyte and fibroblast phenotypes. Keratocytes, isolated from bovine and human corneas, cultured in serum-free medium supplemented with insulin, selenium and transferrin, assumed typical keratocyte morphology, converted to fibroblasts in serum-containing medium and reverted to keratocytes after serum-deprivation. The bovine keratocytes produced spheres under adherent or low attachment conditions, while the human keratocytes produced spheres under low attachment conditions only. The primary keratocytes and fibroblasts expressed vimentin, confirming their mesenchymal origin. Keratocan, considered to be a marker for keratocytes, was also detected in early passage bovine fibroblasts. BMP3 was expressed in keratocytes and keratocyte-derived spheres, while cadherin 5 in keratocytes only, suggesting these as potential keratocyte markers.

Original languageEnglish (US)
Pages (from-to)898-905
Number of pages8
JournalExperimental Eye Research
Volume93
Issue number6
DOIs
StatePublished - Dec 2011

Fingerprint

Corneal Keratocytes
Fibroblasts
Phenotype
Corneal Stroma
Extracellular Matrix
Myofibroblasts
Serum-Free Culture Media
Vimentin
Tissue Engineering
Transferrin
Selenium
Serum
Cornea
Insulin
Wounds and Injuries

Keywords

  • BMP3
  • Cdh5
  • Cornea
  • Fibroblasts
  • Keratocan
  • Keratocyte
  • Lumican
  • Spheres

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Sphere formation from corneal keratocytes and phenotype specific markers. / Scott, Sherri Gae; Jun, Albert; Chakravarti, Shukti.

In: Experimental Eye Research, Vol. 93, No. 6, 12.2011, p. 898-905.

Research output: Contribution to journalArticle

Scott, Sherri Gae ; Jun, Albert ; Chakravarti, Shukti. / Sphere formation from corneal keratocytes and phenotype specific markers. In: Experimental Eye Research. 2011 ; Vol. 93, No. 6. pp. 898-905.
@article{dc0d8379bed2421fb81a5af39ff26471,
title = "Sphere formation from corneal keratocytes and phenotype specific markers",
abstract = "The keratocytes are specialized mesenchymal cells that produce and maintain the extracellular matrix of the corneal stroma. With a typical dendritic and flattened appearance, these cells can morph into fibroblasts and myofibroblasts upon injury, and produce abnormal or fibrotic extracellular matrices detrimental to corneal transparency. Insights into mechanisms that regulate these phenotypic switches and optimal culture conditions that preserve the keratocyte phenotype are important for tissue engineering of the corneal stroma. Like other cell types with self-renewing capacity, keratocytes can form spheres in culture. Here we investigated human and bovine keratocytes with respect to their sphere forming capabilities, and sought to identify potentially distinguishing markers for the keratocyte and fibroblast phenotypes. Keratocytes, isolated from bovine and human corneas, cultured in serum-free medium supplemented with insulin, selenium and transferrin, assumed typical keratocyte morphology, converted to fibroblasts in serum-containing medium and reverted to keratocytes after serum-deprivation. The bovine keratocytes produced spheres under adherent or low attachment conditions, while the human keratocytes produced spheres under low attachment conditions only. The primary keratocytes and fibroblasts expressed vimentin, confirming their mesenchymal origin. Keratocan, considered to be a marker for keratocytes, was also detected in early passage bovine fibroblasts. BMP3 was expressed in keratocytes and keratocyte-derived spheres, while cadherin 5 in keratocytes only, suggesting these as potential keratocyte markers.",
keywords = "BMP3, Cdh5, Cornea, Fibroblasts, Keratocan, Keratocyte, Lumican, Spheres",
author = "Scott, {Sherri Gae} and Albert Jun and Shukti Chakravarti",
year = "2011",
month = "12",
doi = "10.1016/j.exer.2011.10.004",
language = "English (US)",
volume = "93",
pages = "898--905",
journal = "Experimental Eye Research",
issn = "0014-4835",
publisher = "Academic Press Inc.",
number = "6",

}

TY - JOUR

T1 - Sphere formation from corneal keratocytes and phenotype specific markers

AU - Scott, Sherri Gae

AU - Jun, Albert

AU - Chakravarti, Shukti

PY - 2011/12

Y1 - 2011/12

N2 - The keratocytes are specialized mesenchymal cells that produce and maintain the extracellular matrix of the corneal stroma. With a typical dendritic and flattened appearance, these cells can morph into fibroblasts and myofibroblasts upon injury, and produce abnormal or fibrotic extracellular matrices detrimental to corneal transparency. Insights into mechanisms that regulate these phenotypic switches and optimal culture conditions that preserve the keratocyte phenotype are important for tissue engineering of the corneal stroma. Like other cell types with self-renewing capacity, keratocytes can form spheres in culture. Here we investigated human and bovine keratocytes with respect to their sphere forming capabilities, and sought to identify potentially distinguishing markers for the keratocyte and fibroblast phenotypes. Keratocytes, isolated from bovine and human corneas, cultured in serum-free medium supplemented with insulin, selenium and transferrin, assumed typical keratocyte morphology, converted to fibroblasts in serum-containing medium and reverted to keratocytes after serum-deprivation. The bovine keratocytes produced spheres under adherent or low attachment conditions, while the human keratocytes produced spheres under low attachment conditions only. The primary keratocytes and fibroblasts expressed vimentin, confirming their mesenchymal origin. Keratocan, considered to be a marker for keratocytes, was also detected in early passage bovine fibroblasts. BMP3 was expressed in keratocytes and keratocyte-derived spheres, while cadherin 5 in keratocytes only, suggesting these as potential keratocyte markers.

AB - The keratocytes are specialized mesenchymal cells that produce and maintain the extracellular matrix of the corneal stroma. With a typical dendritic and flattened appearance, these cells can morph into fibroblasts and myofibroblasts upon injury, and produce abnormal or fibrotic extracellular matrices detrimental to corneal transparency. Insights into mechanisms that regulate these phenotypic switches and optimal culture conditions that preserve the keratocyte phenotype are important for tissue engineering of the corneal stroma. Like other cell types with self-renewing capacity, keratocytes can form spheres in culture. Here we investigated human and bovine keratocytes with respect to their sphere forming capabilities, and sought to identify potentially distinguishing markers for the keratocyte and fibroblast phenotypes. Keratocytes, isolated from bovine and human corneas, cultured in serum-free medium supplemented with insulin, selenium and transferrin, assumed typical keratocyte morphology, converted to fibroblasts in serum-containing medium and reverted to keratocytes after serum-deprivation. The bovine keratocytes produced spheres under adherent or low attachment conditions, while the human keratocytes produced spheres under low attachment conditions only. The primary keratocytes and fibroblasts expressed vimentin, confirming their mesenchymal origin. Keratocan, considered to be a marker for keratocytes, was also detected in early passage bovine fibroblasts. BMP3 was expressed in keratocytes and keratocyte-derived spheres, while cadherin 5 in keratocytes only, suggesting these as potential keratocyte markers.

KW - BMP3

KW - Cdh5

KW - Cornea

KW - Fibroblasts

KW - Keratocan

KW - Keratocyte

KW - Lumican

KW - Spheres

UR - http://www.scopus.com/inward/record.url?scp=84855922448&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84855922448&partnerID=8YFLogxK

U2 - 10.1016/j.exer.2011.10.004

DO - 10.1016/j.exer.2011.10.004

M3 - Article

C2 - 22032988

AN - SCOPUS:84855922448

VL - 93

SP - 898

EP - 905

JO - Experimental Eye Research

JF - Experimental Eye Research

SN - 0014-4835

IS - 6

ER -