Amniotic fluid promotes the appearance of neural retinal progenitors and neurons in human RPE cell cultures

Retinal pigment epithelial (RPE) cells are capable of differentiating into retinal neurons when induced by the appropriate growth factors. Amniotic fluid contains a variety of growth factors that are crucial for the development of a fetus. In this study, the effects of human amniotic fluid (HAF) on...

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Bibliographic Details
Published in:Molecular vision 2013-11, Vol.19, p.2330-2342
Main Authors: Davari, Maliheh, Soheili, Zahra-Soheila, Ahmadieh, Hamid, Sanie-Jahromi, Fateme, Ghaderi, Shima, Kanavi, Mozhgan Rezaei, Samiei, Shahram, Akrami, Hassan, Haghighi, Massoud, Javidi-Azad, Fahimeh
Format: Article
Language:English
Subjects:
Amniotic Fluid - cytology
Animals
Biomarkers - metabolism
Cattle
Cell Proliferation
Cell Shape
Cells, Cultured
Enzyme-Linked Immunosorbent Assay
Female
Humans
Immunohistochemistry
Neural Stem Cells - cytology
Neural Stem Cells - metabolism
Retinal Neurons - cytology
Retinal Neurons - metabolism
Retinal Pigment Epithelium - cytology
Spheroids, Cellular - cytology
Up-Regulation
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Summary:Retinal pigment epithelial (RPE) cells are capable of differentiating into retinal neurons when induced by the appropriate growth factors. Amniotic fluid contains a variety of growth factors that are crucial for the development of a fetus. In this study, the effects of human amniotic fluid (HAF) on primary RPE cell cultures were evaluated. RPE cells were isolated from the globes of postnatal human cadavers. The isolated cells were plated and grown in DMEM/F12 with 10% fetal bovine serum. To confirm the RPE identity of the cultured cells, they were immunocytochemically examined for the presence of the RPE cell-specific marker RPE65. RPE cultures obtained from passages 2-7 were treated with HAF and examined morphologically for 1 month. To determine whether retinal neurons or progenitors developed in the treated cultures, specific markers for bipolar (protein kinase C isomer α, PKCα), amacrine (cellular retinoic acid-binding protein I, CRABPI), and neural progenitor (NESTIN) cells were sought, and the amount of mRNA was quantified using real-time PCR. Treating RPE cells with HAF led to a significant decrease in the number of RPE65-positive cells, while PKCα- and CRABPI-positive cells were detected in the cultures. Compared with the fetal bovine serum-treated cultures, the levels of mRNAs quantitatively increased by 2-, 20- and 22-fold for NESTIN, PKCα, and CRABPI, respectively. The RPE cultures treated with HAF established spheres containing both pigmented and nonpigmented cells, which expressed neural progenitor markers such as NESTIN. This study showed that HAF can induce RPE cells to transdifferentiate into retinal neurons and progenitor cells, and that it provides a potential source for cell-based therapies to treat retinal diseases.
ISSN:1090-0535