The effect of elevated extracellular glucose on migration, adhesion and proliferation of SV40 transformed human corneal epithelial cells

Alison M. McDermott, Timothy S. Kern, Christopher J Murphy

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Purpose. To examine the effect of elevated extracellular glucose, thus simulating diabetes, on migration, adhesion and proliferation of SV40 transformed human corneal epithelial (HCE) cells. Methods. HCE cells were maintained in serum supplemented media containing 5 mM, 17.5 mM or 38 mM D-glucose. Cell migration was determined using Blind well chambers fitted with fibronectin/collagen I coated filters. In adhesion experiments, cells were allowed to adhere to extracellular matrix protein-coated wells for 90 min at 37°C. Non-adherent cells were removed by washing, then the fluorochrome calcein-AM was added to quantitate the number of attached cells. Proliferation was determined by plating the cells at low density, then quantitating viable cells with calcein-AM 5 to 7 days later. Results. Raising extracellular glucose from 5 mM to 17.5 mM significantly increased cell migration by 42%. When glucose was further raised to 38 mM, migration was not significantly different from that in 5 mM glucose. Adhesion to fibronectin and collagen I (but not IV) was significantly increased (62% and 32% respectively) when cells were cultured in 17.5 mM glucose. Similarly, proliferation was increased by 44%. Adhesion and proliferation tended to be decreased at 38 mM compared to 17.5 mM glucose, but not significantly so. In the presence of 5 mM glucose and mannitol (12.5 mM or 33 mM), neither migration, adhesion nor proliferation were significantly different from that in 5 mM glucose alone. Conclusion. Elevated extracellular glucose modulates migration, adhesion and proliferation of HCE cells. The effects are dependent on the concentration of glucose and are not due to changes in osmolality since mannitol failed to produce similar results. Our in vitro findings suggest that high-glucose effects may directly contribute to the etiology of impaired corneal wound healing in diabetes.

Original languageEnglish (US)
Pages (from-to)924-932
Number of pages9
JournalCurrent Eye Research
Volume17
Issue number9
DOIs
StatePublished - 1998
Externally publishedYes

Fingerprint

Epithelial Cells
Glucose
Mannitol
Fibronectins
Cell Movement
Collagen
Extracellular Matrix Proteins
Fluorescent Dyes
Cell Adhesion
Wound Healing
Osmolar Concentration
Cultured Cells
Cell Count
Serum

Keywords

  • Adhesion
  • Corneal epithelium
  • Diabetes
  • Human
  • Migration
  • Proliferation

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

The effect of elevated extracellular glucose on migration, adhesion and proliferation of SV40 transformed human corneal epithelial cells. / McDermott, Alison M.; Kern, Timothy S.; Murphy, Christopher J.

In: Current Eye Research, Vol. 17, No. 9, 1998, p. 924-932.

Research output: Contribution to journalArticle

@article{f3bbb17e00e34e5aabebcc4f0e73dfe7,
title = "The effect of elevated extracellular glucose on migration, adhesion and proliferation of SV40 transformed human corneal epithelial cells",
abstract = "Purpose. To examine the effect of elevated extracellular glucose, thus simulating diabetes, on migration, adhesion and proliferation of SV40 transformed human corneal epithelial (HCE) cells. Methods. HCE cells were maintained in serum supplemented media containing 5 mM, 17.5 mM or 38 mM D-glucose. Cell migration was determined using Blind well chambers fitted with fibronectin/collagen I coated filters. In adhesion experiments, cells were allowed to adhere to extracellular matrix protein-coated wells for 90 min at 37°C. Non-adherent cells were removed by washing, then the fluorochrome calcein-AM was added to quantitate the number of attached cells. Proliferation was determined by plating the cells at low density, then quantitating viable cells with calcein-AM 5 to 7 days later. Results. Raising extracellular glucose from 5 mM to 17.5 mM significantly increased cell migration by 42{\%}. When glucose was further raised to 38 mM, migration was not significantly different from that in 5 mM glucose. Adhesion to fibronectin and collagen I (but not IV) was significantly increased (62{\%} and 32{\%} respectively) when cells were cultured in 17.5 mM glucose. Similarly, proliferation was increased by 44{\%}. Adhesion and proliferation tended to be decreased at 38 mM compared to 17.5 mM glucose, but not significantly so. In the presence of 5 mM glucose and mannitol (12.5 mM or 33 mM), neither migration, adhesion nor proliferation were significantly different from that in 5 mM glucose alone. Conclusion. Elevated extracellular glucose modulates migration, adhesion and proliferation of HCE cells. The effects are dependent on the concentration of glucose and are not due to changes in osmolality since mannitol failed to produce similar results. Our in vitro findings suggest that high-glucose effects may directly contribute to the etiology of impaired corneal wound healing in diabetes.",
keywords = "Adhesion, Corneal epithelium, Diabetes, Human, Migration, Proliferation",
author = "McDermott, {Alison M.} and Kern, {Timothy S.} and Murphy, {Christopher J}",
year = "1998",
doi = "10.1076/ceyr.17.9.924.5133",
language = "English (US)",
volume = "17",
pages = "924--932",
journal = "Current Eye Research",
issn = "0271-3683",
publisher = "Informa Healthcare",
number = "9",

}

TY - JOUR

T1 - The effect of elevated extracellular glucose on migration, adhesion and proliferation of SV40 transformed human corneal epithelial cells

AU - McDermott, Alison M.

AU - Kern, Timothy S.

AU - Murphy, Christopher J

PY - 1998

Y1 - 1998

N2 - Purpose. To examine the effect of elevated extracellular glucose, thus simulating diabetes, on migration, adhesion and proliferation of SV40 transformed human corneal epithelial (HCE) cells. Methods. HCE cells were maintained in serum supplemented media containing 5 mM, 17.5 mM or 38 mM D-glucose. Cell migration was determined using Blind well chambers fitted with fibronectin/collagen I coated filters. In adhesion experiments, cells were allowed to adhere to extracellular matrix protein-coated wells for 90 min at 37°C. Non-adherent cells were removed by washing, then the fluorochrome calcein-AM was added to quantitate the number of attached cells. Proliferation was determined by plating the cells at low density, then quantitating viable cells with calcein-AM 5 to 7 days later. Results. Raising extracellular glucose from 5 mM to 17.5 mM significantly increased cell migration by 42%. When glucose was further raised to 38 mM, migration was not significantly different from that in 5 mM glucose. Adhesion to fibronectin and collagen I (but not IV) was significantly increased (62% and 32% respectively) when cells were cultured in 17.5 mM glucose. Similarly, proliferation was increased by 44%. Adhesion and proliferation tended to be decreased at 38 mM compared to 17.5 mM glucose, but not significantly so. In the presence of 5 mM glucose and mannitol (12.5 mM or 33 mM), neither migration, adhesion nor proliferation were significantly different from that in 5 mM glucose alone. Conclusion. Elevated extracellular glucose modulates migration, adhesion and proliferation of HCE cells. The effects are dependent on the concentration of glucose and are not due to changes in osmolality since mannitol failed to produce similar results. Our in vitro findings suggest that high-glucose effects may directly contribute to the etiology of impaired corneal wound healing in diabetes.

AB - Purpose. To examine the effect of elevated extracellular glucose, thus simulating diabetes, on migration, adhesion and proliferation of SV40 transformed human corneal epithelial (HCE) cells. Methods. HCE cells were maintained in serum supplemented media containing 5 mM, 17.5 mM or 38 mM D-glucose. Cell migration was determined using Blind well chambers fitted with fibronectin/collagen I coated filters. In adhesion experiments, cells were allowed to adhere to extracellular matrix protein-coated wells for 90 min at 37°C. Non-adherent cells were removed by washing, then the fluorochrome calcein-AM was added to quantitate the number of attached cells. Proliferation was determined by plating the cells at low density, then quantitating viable cells with calcein-AM 5 to 7 days later. Results. Raising extracellular glucose from 5 mM to 17.5 mM significantly increased cell migration by 42%. When glucose was further raised to 38 mM, migration was not significantly different from that in 5 mM glucose. Adhesion to fibronectin and collagen I (but not IV) was significantly increased (62% and 32% respectively) when cells were cultured in 17.5 mM glucose. Similarly, proliferation was increased by 44%. Adhesion and proliferation tended to be decreased at 38 mM compared to 17.5 mM glucose, but not significantly so. In the presence of 5 mM glucose and mannitol (12.5 mM or 33 mM), neither migration, adhesion nor proliferation were significantly different from that in 5 mM glucose alone. Conclusion. Elevated extracellular glucose modulates migration, adhesion and proliferation of HCE cells. The effects are dependent on the concentration of glucose and are not due to changes in osmolality since mannitol failed to produce similar results. Our in vitro findings suggest that high-glucose effects may directly contribute to the etiology of impaired corneal wound healing in diabetes.

KW - Adhesion

KW - Corneal epithelium

KW - Diabetes

KW - Human

KW - Migration

KW - Proliferation

UR - http://www.scopus.com/inward/record.url?scp=0031654517&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031654517&partnerID=8YFLogxK

U2 - 10.1076/ceyr.17.9.924.5133

DO - 10.1076/ceyr.17.9.924.5133

M3 - Article

C2 - 9746440

AN - SCOPUS:0031654517

VL - 17

SP - 924

EP - 932

JO - Current Eye Research

JF - Current Eye Research

SN - 0271-3683

IS - 9

ER -