Generation of Retinal Pigmented Epithelium-Like Cells from Pigmented Spheres Differentiated from Bone Marrow Stromal Cell-Derived Neurospheres
10.1007/s13770-019-00183-1
- Author:
Hamid Aboutaleb KADKHODAEIAN
1
;
Taki TIRAIHI
;
Hamid AHMADIEH
;
Hossein ZIAEI
;
Narsis DAFTARIAN
;
Taher TAHERI
Author Information
1. Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 3519899951, 3514799422 Tehran, Iran. takialtr@modares.ac.ir, ttiraihi@gmail.com
- Publication Type:Original Article
- Keywords:
Bone marrow stromal stem cells;
Neurosphere;
Retinal pigment epithelium;
Transdifferentiation
- MeSH:
Animals;
Blindness;
Bone Marrow;
Epithelium;
Fibronectins;
Gene Expression;
Genes, Homeobox;
Humans;
Immunohistochemistry;
Male;
Nestin;
Phagocytes;
Phagocytosis;
Rats;
Retinal Degeneration;
Retinal Pigment Epithelium;
Retinaldehyde;
Retinitis Pigmentosa;
Stem Cells
- From:
Tissue Engineering and Regenerative Medicine
2019;16(3):253-263
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: Retinal degeneration causes blindness, and cell replacement is a potential therapy. The purpose of this study is to formation of pigmented neurospheres in a simple medium, low-cost, high-performance manner over a short period of time while expressing markers of RPE cells and the activation of specific genes of the pigment cells. Also, these neurospheres have the ability to produce a monolayer of retinal pigment epithelium-like cells (RPELC) with the ability of photoreceptor outer segment phagocytosis. METHODS: BMSC were isolated from pigmented hooded male rats and were immunoreactive to BMSC markers, then converted into neurospheres, differentiated into pigmented spheres (PS), and characterized using Retinal pigment epithelium-specific 65 kDa protein (RPE65), Retinaldehyde-binding protein 1 (CRALBP) and orthodenticle homeobox 2 (OTX2) markers by immunocytochemistry, RT-PCR and RT-qPCR. The PS were harvested into RPELC. The functionality of RPELC was evaluated by phagocytosis of fluorescein-labeled photoreceptor outer segment. RESULTS: The BMSC immunophenotype was confirmed by immunostained for fibronectin, CD90, CD166 and CD44. These cells differentiated into osteogenic and lipogenic cells. The generated neurospheres were immunoreactive to nestin and stemness genes. The PS after 7–14 days were positive for RPE65 (92.76–100%), CRALBP (95.21–100%) and OTX2 (94.88–100%), and after 30 days RT-PCR, qPCR revealed increasing in gene expression. The PS formed a single layer of RPELC after cultivation and phagocyte photoreceptor outer segments. CONCLUSION: Bone marrow stromal stem cells can differentiate into functional retinal pigmented epithelium cells in a simple, low-cost, high-performancemanner over a short period of time. These cells due to expressing theRPELCgenes andmarkers can be used in cell replacement therapy for degenerative diseases including age-relatedmacular degeneration as well as retinitis pigmentosa.
