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Secretion Profile of Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium During Wound Healing.
Invest Ophthalmol Vis Sci. 2016 Aug 01; 57(10):4428-4441.IO

Abstract

Purpose

The purpose of this study was to characterize the secretion profile of induced pluripotent stem cell-derived retinal pigment epithelium (iPS-RPE) during wound healing. iPS-RPE was used to develop an in vitro wound healing model. We hypothesized that iPS-RPE secretes cytokines and growth factors which act in an autocrine manner to promote migration and proliferation of cells during wound healing.

Methods

iPS-RPE was grown in transwells until fully confluent and pigmented. The monolayers were scratched to induce a wound. Levels of Ki-67, β-catenin, e-cadherin, n-cadherin, and S100A4 expression were analyzed by immunofluorescent labeling. Cell culture medium samples were collected from both the apical and basolateral sides of the transwells every 72 hours for 21 days. The medium samples were analyzed using multiplex ELISA to detect secreted growth factors and cytokines. The effects of conditioned medium on collagen gel contraction, cell proliferation, and migration were measured.

Results

iPS-RPE underwent epithelial-mesenchymal transition (EMT) during wound healing as indicated by the translocation of β-catenin to the nucleus, cadherin switch, and expression of S100A4. GRO, GM-CSF, MCP-1, IL-6, and IL-8 were secreted by both the control and the wounded cell cultures. VEGF, FGF-2, and TGFβ expression were detected at higher levels after wounding than those in control. The proteins were found to be secreted in a polarized manner. The conditioned medium from wounded monolayers promoted collagen gel contraction, as well as proliferation and migration of ARPE 19 cells.

Conclusions

These results indicate that after the monolayer is wounded, iPS-RPE secretes proteins into the culture medium that promote increased proliferation, contraction, and migration.

Authors+Show Affiliations

Ocular Trauma Task Area US Army Institute of Surgical Research, Joint Base San Antonio-Fort Sam Houston, Texas, United States.Ocular Trauma Task Area US Army Institute of Surgical Research, Joint Base San Antonio-Fort Sam Houston, Texas, United States.Ocular Trauma Task Area US Army Institute of Surgical Research, Joint Base San Antonio-Fort Sam Houston, Texas, United States.Ocular Trauma Task Area US Army Institute of Surgical Research, Joint Base San Antonio-Fort Sam Houston, Texas, United States.Ocular Trauma Task Area US Army Institute of Surgical Research, Joint Base San Antonio-Fort Sam Houston, Texas, United States.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27750286

Citation

Greene, Whitney A., et al. "Secretion Profile of Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium During Wound Healing." Investigative Ophthalmology & Visual Science, vol. 57, no. 10, 2016, pp. 4428-4441.
Greene WA, Burke TA, Por ED, et al. Secretion Profile of Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium During Wound Healing. Invest Ophthalmol Vis Sci. 2016;57(10):4428-4441.
Greene, W. A., Burke, T. A., Por, E. D., Kaini, R. R., & Wang, H. C. (2016). Secretion Profile of Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium During Wound Healing. Investigative Ophthalmology & Visual Science, 57(10), 4428-4441. https://doi.org/10.1167/iovs.16-19192
Greene WA, et al. Secretion Profile of Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium During Wound Healing. Invest Ophthalmol Vis Sci. 2016 Aug 1;57(10):4428-4441. PubMed PMID: 27750286.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Secretion Profile of Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium During Wound Healing. AU - Greene,Whitney A, AU - Burke,Teresa A, AU - Por,Elaine D, AU - Kaini,Ramesh R, AU - Wang,Heuy-Ching, PY - 2016/10/18/pubmed PY - 2017/5/23/medline PY - 2016/10/18/entrez SP - 4428 EP - 4441 JF - Investigative ophthalmology & visual science JO - Invest Ophthalmol Vis Sci VL - 57 IS - 10 N2 - Purpose: The purpose of this study was to characterize the secretion profile of induced pluripotent stem cell-derived retinal pigment epithelium (iPS-RPE) during wound healing. iPS-RPE was used to develop an in vitro wound healing model. We hypothesized that iPS-RPE secretes cytokines and growth factors which act in an autocrine manner to promote migration and proliferation of cells during wound healing. Methods: iPS-RPE was grown in transwells until fully confluent and pigmented. The monolayers were scratched to induce a wound. Levels of Ki-67, β-catenin, e-cadherin, n-cadherin, and S100A4 expression were analyzed by immunofluorescent labeling. Cell culture medium samples were collected from both the apical and basolateral sides of the transwells every 72 hours for 21 days. The medium samples were analyzed using multiplex ELISA to detect secreted growth factors and cytokines. The effects of conditioned medium on collagen gel contraction, cell proliferation, and migration were measured. Results: iPS-RPE underwent epithelial-mesenchymal transition (EMT) during wound healing as indicated by the translocation of β-catenin to the nucleus, cadherin switch, and expression of S100A4. GRO, GM-CSF, MCP-1, IL-6, and IL-8 were secreted by both the control and the wounded cell cultures. VEGF, FGF-2, and TGFβ expression were detected at higher levels after wounding than those in control. The proteins were found to be secreted in a polarized manner. The conditioned medium from wounded monolayers promoted collagen gel contraction, as well as proliferation and migration of ARPE 19 cells. Conclusions: These results indicate that after the monolayer is wounded, iPS-RPE secretes proteins into the culture medium that promote increased proliferation, contraction, and migration. SN - 1552-5783 UR - https://www.unboundmedicine.com/medline/citation/27750286/Secretion_Profile_of_Induced_Pluripotent_Stem_Cell_Derived_Retinal_Pigment_Epithelium_During_Wound_Healing_ L2 - https://iovs.arvojournals.org/article.aspx?doi=10.1167/iovs.16-19192 DB - PRIME DP - Unbound Medicine ER -