Bone marrow-derived stem cells differentiate into retinal pigment epithelium-like cells in vitro but are not able to repair retinal degeneration in vivo

Purpose of study: Neurosphere-free trans-differentiation of bone marrow stem cells (NFT-BMSCs) into retinal pigment epithelium (RPE) and retinal cells (RCs) in vitro could offer a unique opportunity for the study of cell-replacement in degenerative eye diseases. In this respect, a simple, and efficient protocol for getting retinal cells from trans-differentiation of rat BMSCs in the neurosphere-free state are reported. Methods: Extracted BMSCs from hooded pigment rats were exposed to a single-step protocol, including neurosphere-free, containing a cocktail medium which induced trans-differentiation into retinal pigment epithelium (RPE) and retinal cells. Results: The results showed morphological differentiation changes in vitro. Also, expressed retinal pigment epithelium and retinal cell markers such as Otx2 (23.45%), RPE65 (91.54%), CRALBP (91.21%), VEGF (94.79%), Rhodopsin (57.19%), GFAP (28.33%), and NF200 (24.55%). Conclusion: Overall, these findings showed a protocol, without using bFGF, EGF, and B-27 supplement which makes it possible to obtain RPE and retinal cells in vitro.

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In Vitro Differentiation of Human Bone Marrow Stem Cells into Retinal Pigment Epithelium

American Journal of Bioscience and Bioengineering ◽

10.11648/j.bio.s.2015030401.18 ◽

2015 ◽

Vol 3 (4) ◽

pp. 51

Author(s):

Nehal Draz

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Retinal Pigment Epithelium ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Bone Marrow Stem Cells ◽

Human Bone ◽

Human Bone Marrow ◽

In Vitro Differentiation

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Inhibition of DUSP6 Activates Autophagy and Rescues the Retinal Pigment Epithelium in Sodium Iodate-Induced Retinal Degeneration Models In Vivo and In Vitro

Biomedicines ◽

10.3390/biomedicines10010159 ◽

2022 ◽

Vol 10 (1) ◽

pp. 159

Author(s):

Hao-Yu Tsai ◽

Henkie Isahwan Ahmad Mulyadi Lai ◽

Zhang-Yuan Chen ◽

Tai-Chi Lin ◽

Winnie Khor ◽

...

Keyword(s):

Retinal Pigment Epithelium ◽

Retinal Degeneration ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Autophagic Flux ◽

Erk Pathway ◽

Sodium Iodate ◽

Autophagy Flux

Autophagy plays a protective role in the retinal pigment epithelium (RPE) by eliminating damaged organelles in response to reactive oxygen species (ROS). Dual-specificity protein phosphatase 6 (DUSP6), which belongs to the DUSP subfamily, works as a negative-feedback regulator of the extracellular signal-regulated kinase (ERK) pathway. However, the complex interplay between DUSP6 and autophagy induced by ROS in RPE is yet to be investigated. To investigate the relationship between DUSP6 and autophagy, we exposed the ARPE-19 cell line and C57BL/6N mice to sodium iodate (NaIO3) as an oxidative stress inducer. Our data showed that the inhibition of DUSP6 activity promotes autophagy flux through the ERK pathway via the upregulation of immunoblotting expression in ARPE-19 cells. Live imaging showed a significant increase in autophagic flux activities, which suggested the restoration autophagy after treatment with the DUSP6 inhibitor. Furthermore, the mouse RPE layer exhibited an irregular structure and abnormal deposits following NaIO3 injection. The retina layer was recovered after being treated with DUSP6 inhibitor; this suggests that DUSP6 inhibitor can rescue retinal damage by restoring the mouse retina’s autophagy flux. This study suggests that the upregulation of DUSP6 can cause autophagy flux malfunctions in the RPE. The DUSP6 inhibitor can restore autophagy induction, which may serve as a potential therapeutic approach for retinal degeneration disease.

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The cytoskeletal elements of human retinal pigment epithelium: in vitro and in vivo

Journal of Cell Science ◽

10.1242/jcs.91.2.303 ◽

1988 ◽

Vol 91 (2) ◽

pp. 303-312

Author(s):

N.M. McKechnie ◽

M. Boulton ◽

H.L. Robey ◽

F.J. Savage ◽

I. Grierson

Keyword(s):

Retinal Pigment Epithelium ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Cytokeratin 18 ◽

Rpe Cells ◽

Human Retinal Pigment Epithelium ◽

Cytoskeletal Elements

The cytoskeletal elements of normal (in situ) and cultured human retinal pigment epithelium (RPE) were studied by a variety of immunocytochemical techniques. Primary antibodies to vimentin and cytokeratins were used. Positive immunoreactivity for vimentin was obtained with in situ and cultured material. The pattern of reactivity obtained with antisera and monoclonals to cytokeratins was more complex. Cytokeratin immunoreactivity could be demonstrated in situ and in cultured cells. The pattern of cytokeratin expression was similar to that of simple or glandular epithelia. A monoclonal antibody that specifically recognizes cytokeratin 18 identified a population of cultured RPE cells that had particularly well-defined filamentous networks within their cytoplasm. Freshly isolated RPE was cytokeratin 18 negative by immunofluorescence, but upon culture cytokeratin 18 positive cells were identifiable. Cytokeratin 18 positive cells were identified in all RPE cultures (other than early primaries), regardless of passage number, age or sex of the donor. In post-confluent cultures cytokeratin 18 cells were identified growing over cytokeratin 18 negative cells, suggesting an association of cytokeratin 18 immunoreactivity with cell proliferation. Immunofluorescence studies of retinal scar tissue from two individuals revealed the presence of numerous cytokeratin 18 positive cells. These findings indicate that RPE cells can be identified by their cytokeratin immunoreactivity and that the overt expression of cytokeratin 18 may be associated with proliferation of human RPE both in vitro and in vivo.

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JAM-C maintains VEGR2 expression to promote retinal pigment epithelium cell survival under oxidative stress

Thrombosis and Haemostasis ◽

10.1160/th16-11-0885 ◽

2017 ◽

Vol 117 (04) ◽

pp. 750-757

Author(s):

Xin Jia ◽

Chen Zhao ◽

Qishan Chen ◽

Yuxiang Du ◽

Lijuan Huang ◽

...

Keyword(s):

Oxidative Stress ◽

Retinal Pigment Epithelium ◽

Cell Survival ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Retinal Pigment Epithelium Cell ◽

Photoreceptor Cells ◽

Rpe Cells

SummaryJunctional adhesion molecule-C (JAM-C) has been shown to play critical roles during development and in immune responses. However, its role in adult eyes under oxidative stress remains poorly understood. Here, we report that JAM-C is abundantly expressed in adult mouse retinae and choroids in vivo and in cultured retinal pigment epithelium (RPE) and photoreceptor cells in vitro. Importantly, both JAM-C expression and its membrane localisation are downregulated by H2O2-induced oxidative stress. Under H2O2-induced oxidative stress, JAM-C is critically required for the survival of human RPE cells. Indeed, loss of JAM-C by siRNA knockdown decreased RPE cell survival. Mechanistically, we show that JAM-C is required to maintain VEGFR2 expression in RPE cells, and VEGFR2 plays an important role in keeping the RPE cells viable since overexpression of VEGFR2 partially restored impaired RPE survival caused by JAM-C knockdown and increased RPE survival. We further show that JAM-C regulates VEGFR2 expression and, in turn, modulates p38 phosphorylation. Together, our data demonstrate that JAM-C plays an important role in maintaining VEGR2 expression to promote RPE cell survival under oxidative stress. Given the vital importance of RPE in the eye, approaches that can modulate JAM-C expression may have therapeutic values in treating diseases with impaired RPE survival.

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Stem Cells are Mobilized from the Bone Marrow into the Peripheral Circulation in Response to Retinal Pigment Epithelium Damage—A Pathophysiological Attempt to Induce Endogenous Regeneration

Current Eye Research ◽

10.3109/02713683.2011.576796 ◽

2011 ◽

Vol 36 (7) ◽

pp. 663-672 ◽

Cited By ~ 15

Author(s):

Anna Machalińska ◽

Patrycja Kłos ◽

Bartłomiej Baumert ◽

Magdalena Baśkiewicz ◽

Miłosz Kawa ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Retinal Pigment Epithelium ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Peripheral Circulation

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Clinical-grade stem cell–derived retinal pigment epithelium patch rescues retinal degeneration in rodents and pigs

Science Translational Medicine ◽

10.1126/scitranslmed.aat5580 ◽

2019 ◽

Vol 11 (475) ◽

pp. eaat5580 ◽

Cited By ~ 57

Author(s):

Ruchi Sharma ◽

Vladimir Khristov ◽

Aaron Rising ◽

Balendu Shekhar Jha ◽

Roba Dejene ◽

...

Keyword(s):

Stem Cell ◽

Retinal Pigment Epithelium ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Age Related Macular Degeneration ◽

Clinical Grade ◽

Functional Validation ◽

Age Related

Considerable progress has been made in testing stem cell–derived retinal pigment epithelium (RPE) as a potential therapy for age-related macular degeneration (AMD). However, the recent reports of oncogenic mutations in induced pluripotent stem cells (iPSCs) underlie the need for robust manufacturing and functional validation of clinical-grade iPSC-derived RPE before transplantation. Here, we developed oncogenic mutation-free clinical-grade iPSCs from three AMD patients and differentiated them into clinical-grade iPSC-RPE patches on biodegradable scaffolds. Functional validation of clinical-grade iPSC-RPE patches revealed specific features that distinguished transplantable from nontransplantable patches. Compared to RPE cells in suspension, our biodegradable scaffold approach improved integration and functionality of RPE patches in rats and in a porcine laser-induced RPE injury model that mimics AMD-like eye conditions. Our results suggest that the in vitro and in vivo preclinical functional validation of iPSC-RPE patches developed here might ultimately be useful for evaluation and optimization of autologous iPSC-based therapies.

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