Cell-deposited matrix improves retinal pigment epithelium survival on aged submacular human Bruch's membrane

Ilene K. Sugino, Vamsi K. Gullapalli, Qian Sun, Jianqiu Wang, Celia F. Nunes, Noounanong Cheewatrakoolpong, Adam C. Johnson, Benjamin C. Degner, Jianyuan Hua, Tong Liu, Wei Chen, Hong Li, Marco A. Zarbin

Research output: Contribution to journalArticle

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Abstract

PURPOSE. To determine whether resurfacing submacular human Bruch's membrane with a cell-deposited extracellular matrix (ECM) improves retinal pigment epithelial (RPE) survival. METHODS. Bovine corneal endothelial (BCE) cells were seeded onto the inner collagenous layer of submacular Bruch's membrane explants of human donor eyes to allow ECM deposition. Control explants from fellow eyes were cultured in medium only. The deposited ECM was exposed by removing BCE. Fetal RPE cells were then cultured on these explants for 1, 14, or 21 days. The explants were analyzed quantitatively by light microscopy and scanning electron microscopy. Surviving RPE cells from explants cultured for 21 days were harvested to compare bestrophin and RPE65 mRNA expression. Mass spectroscopy was performed on BCE-ECM to examine the protein composition. RESULTS. The BCE-treated explants showed significantly higher RPE nuclear density than did the control explants at all time points. RPE expressed more differentiated features on BCEtreated explants than on untreated explants, but expressed very little mRNA for bestrophin or RPE65. The untreated young (50 years) and African American submacular Bruch's membrane explants supported significantly higher RPE nuclear densities (NDs) than did the Caucasian explants. These differences were reduced or nonexistent in the BCE-ECM-treated explants. Proteins identified in the BCE-ECM included ECM proteins, ECM-associated proteins, cell membrane proteins, and intracellular proteins. CONCLUSIONS. Increased RPE survival can be achieved on aged submacular human Bruch's membrane by resurfacing the latter with a cell-deposited ECM. Caucasian eyes seem to benefit the most, as cell survival is the worst on submacular Bruch's membrane in these eyes.

Original languageEnglish (US)
Pages (from-to)1345-1358
Number of pages14
JournalInvestigative Ophthalmology and Visual Science
Volume52
Issue number3
DOIs
StatePublished - Mar 1 2011

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Bruch Membrane
Retinal Pigments
Retinal Pigment Epithelium
Extracellular Matrix
Extracellular Matrix Proteins
Epithelial Cells
Messenger RNA
Proteins
Survival
African Americans
Electron Scanning Microscopy
Microscopy
Mass Spectrometry
Cell Survival
Membrane Proteins
Endothelial Cells
Tissue Donors
Light

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Sugino, Ilene K. ; Gullapalli, Vamsi K. ; Sun, Qian ; Wang, Jianqiu ; Nunes, Celia F. ; Cheewatrakoolpong, Noounanong ; Johnson, Adam C. ; Degner, Benjamin C. ; Hua, Jianyuan ; Liu, Tong ; Chen, Wei ; Li, Hong ; Zarbin, Marco A. / Cell-deposited matrix improves retinal pigment epithelium survival on aged submacular human Bruch's membrane. In: Investigative Ophthalmology and Visual Science. 2011 ; Vol. 52, No. 3. pp. 1345-1358.
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abstract = "PURPOSE. To determine whether resurfacing submacular human Bruch's membrane with a cell-deposited extracellular matrix (ECM) improves retinal pigment epithelial (RPE) survival. METHODS. Bovine corneal endothelial (BCE) cells were seeded onto the inner collagenous layer of submacular Bruch's membrane explants of human donor eyes to allow ECM deposition. Control explants from fellow eyes were cultured in medium only. The deposited ECM was exposed by removing BCE. Fetal RPE cells were then cultured on these explants for 1, 14, or 21 days. The explants were analyzed quantitatively by light microscopy and scanning electron microscopy. Surviving RPE cells from explants cultured for 21 days were harvested to compare bestrophin and RPE65 mRNA expression. Mass spectroscopy was performed on BCE-ECM to examine the protein composition. RESULTS. The BCE-treated explants showed significantly higher RPE nuclear density than did the control explants at all time points. RPE expressed more differentiated features on BCEtreated explants than on untreated explants, but expressed very little mRNA for bestrophin or RPE65. The untreated young (50 years) and African American submacular Bruch's membrane explants supported significantly higher RPE nuclear densities (NDs) than did the Caucasian explants. These differences were reduced or nonexistent in the BCE-ECM-treated explants. Proteins identified in the BCE-ECM included ECM proteins, ECM-associated proteins, cell membrane proteins, and intracellular proteins. CONCLUSIONS. Increased RPE survival can be achieved on aged submacular human Bruch's membrane by resurfacing the latter with a cell-deposited ECM. Caucasian eyes seem to benefit the most, as cell survival is the worst on submacular Bruch's membrane in these eyes.",
author = "Sugino, {Ilene K.} and Gullapalli, {Vamsi K.} and Qian Sun and Jianqiu Wang and Nunes, {Celia F.} and Noounanong Cheewatrakoolpong and Johnson, {Adam C.} and Degner, {Benjamin C.} and Jianyuan Hua and Tong Liu and Wei Chen and Hong Li and Zarbin, {Marco A.}",
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Sugino, IK, Gullapalli, VK, Sun, Q, Wang, J, Nunes, CF, Cheewatrakoolpong, N, Johnson, AC, Degner, BC, Hua, J, Liu, T, Chen, W, Li, H & Zarbin, MA 2011, 'Cell-deposited matrix improves retinal pigment epithelium survival on aged submacular human Bruch's membrane', Investigative Ophthalmology and Visual Science, vol. 52, no. 3, pp. 1345-1358. https://doi.org/10.1167/iovs.10-6112

Cell-deposited matrix improves retinal pigment epithelium survival on aged submacular human Bruch's membrane. / Sugino, Ilene K.; Gullapalli, Vamsi K.; Sun, Qian; Wang, Jianqiu; Nunes, Celia F.; Cheewatrakoolpong, Noounanong; Johnson, Adam C.; Degner, Benjamin C.; Hua, Jianyuan; Liu, Tong; Chen, Wei; Li, Hong; Zarbin, Marco A.

In: Investigative Ophthalmology and Visual Science, Vol. 52, No. 3, 01.03.2011, p. 1345-1358.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cell-deposited matrix improves retinal pigment epithelium survival on aged submacular human Bruch's membrane

AU - Sugino, Ilene K.

AU - Gullapalli, Vamsi K.

AU - Sun, Qian

AU - Wang, Jianqiu

AU - Nunes, Celia F.

AU - Cheewatrakoolpong, Noounanong

AU - Johnson, Adam C.

AU - Degner, Benjamin C.

AU - Hua, Jianyuan

AU - Liu, Tong

AU - Chen, Wei

AU - Li, Hong

AU - Zarbin, Marco A.

PY - 2011/3/1

Y1 - 2011/3/1

N2 - PURPOSE. To determine whether resurfacing submacular human Bruch's membrane with a cell-deposited extracellular matrix (ECM) improves retinal pigment epithelial (RPE) survival. METHODS. Bovine corneal endothelial (BCE) cells were seeded onto the inner collagenous layer of submacular Bruch's membrane explants of human donor eyes to allow ECM deposition. Control explants from fellow eyes were cultured in medium only. The deposited ECM was exposed by removing BCE. Fetal RPE cells were then cultured on these explants for 1, 14, or 21 days. The explants were analyzed quantitatively by light microscopy and scanning electron microscopy. Surviving RPE cells from explants cultured for 21 days were harvested to compare bestrophin and RPE65 mRNA expression. Mass spectroscopy was performed on BCE-ECM to examine the protein composition. RESULTS. The BCE-treated explants showed significantly higher RPE nuclear density than did the control explants at all time points. RPE expressed more differentiated features on BCEtreated explants than on untreated explants, but expressed very little mRNA for bestrophin or RPE65. The untreated young (50 years) and African American submacular Bruch's membrane explants supported significantly higher RPE nuclear densities (NDs) than did the Caucasian explants. These differences were reduced or nonexistent in the BCE-ECM-treated explants. Proteins identified in the BCE-ECM included ECM proteins, ECM-associated proteins, cell membrane proteins, and intracellular proteins. CONCLUSIONS. Increased RPE survival can be achieved on aged submacular human Bruch's membrane by resurfacing the latter with a cell-deposited ECM. Caucasian eyes seem to benefit the most, as cell survival is the worst on submacular Bruch's membrane in these eyes.

AB - PURPOSE. To determine whether resurfacing submacular human Bruch's membrane with a cell-deposited extracellular matrix (ECM) improves retinal pigment epithelial (RPE) survival. METHODS. Bovine corneal endothelial (BCE) cells were seeded onto the inner collagenous layer of submacular Bruch's membrane explants of human donor eyes to allow ECM deposition. Control explants from fellow eyes were cultured in medium only. The deposited ECM was exposed by removing BCE. Fetal RPE cells were then cultured on these explants for 1, 14, or 21 days. The explants were analyzed quantitatively by light microscopy and scanning electron microscopy. Surviving RPE cells from explants cultured for 21 days were harvested to compare bestrophin and RPE65 mRNA expression. Mass spectroscopy was performed on BCE-ECM to examine the protein composition. RESULTS. The BCE-treated explants showed significantly higher RPE nuclear density than did the control explants at all time points. RPE expressed more differentiated features on BCEtreated explants than on untreated explants, but expressed very little mRNA for bestrophin or RPE65. The untreated young (50 years) and African American submacular Bruch's membrane explants supported significantly higher RPE nuclear densities (NDs) than did the Caucasian explants. These differences were reduced or nonexistent in the BCE-ECM-treated explants. Proteins identified in the BCE-ECM included ECM proteins, ECM-associated proteins, cell membrane proteins, and intracellular proteins. CONCLUSIONS. Increased RPE survival can be achieved on aged submacular human Bruch's membrane by resurfacing the latter with a cell-deposited ECM. Caucasian eyes seem to benefit the most, as cell survival is the worst on submacular Bruch's membrane in these eyes.

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U2 - 10.1167/iovs.10-6112

DO - 10.1167/iovs.10-6112

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