Matrigel and Activin A promote cell-cell contact and anti-apoptotic activity in cultured human retinal pigment epithelium cells

Guo, Xiaoling; Zhu, Deliang; Lian, Ruiling; Han, Yuting; Guo, Yanjie; Li, Zhijie; Tang, Shibo; Chen, Jiansu

doi:10.1016/j.exer.2016.04.021

Cited by 12 publications

(12 citation statements)

References 47 publications

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“…The physical properties required include biostability, porosity and suitable mechanical strength for surgical handling. It is well known that the surface properties of the underlying substrate directly influences the cells’ ability to form a differentiated monolayer [8]. It is highly likely that the production of a stable basement membrane by RPE cells grown on a synthetic membrane will be crucial to the long-term behaviour of the transplanted construct.…”

Section: Introductionmentioning

confidence: 99%

The formation of a functional retinal pigment epithelium occurs on porous polytetrafluoroethylene substrates independently of the surface chemistry

Kearns

Tasker

Zhuola

et al. 2017

J Mater Sci: Mater Med

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Subretinal transplantation of functioning retinal pigment epithelial (RPE) cells may have the potential to preserve or restore vision in patients affected by blinding diseases such as age-related macular degeneration (AMD). One of the critical steps in achieving this is the ability to grow a functioning retinal pigment epithelium, which may need a substrate on which to grow and to aid transplantation. Tailoring the physical and chemical properties of the substrate should help the engineered tissue to function in the long term. The purpose of the study was to determine whether a functioning monolayer of RPE cells could be produced on expanded polytetrafluoroethylene substrates modified by either an ammonia plasma treatment or an n-Heptylamine coating, and whether the difference in surface chemistries altered the extracellular matrix the cells produced. Primary human RPE cells were able to form a functional, cobblestone monolayer on both substrates, but the formation of an extracellular matrix to exhibit a network structure took months, whereas on non-porous substrates with the same surface chemistry, a similar appearance was observed after a few weeks. This study suggests that the surface chemistry of these materials may not be the most critical factor in the development of growth of a functional monolayer of RPE cells as long as the cells can attach and proliferate on the surface. This has important implications in the design of strategies to optimise the clinical outcomes of subretinal transplant procedures.Graphical Abstract

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Section: Introductionmentioning

confidence: 99%

The formation of a functional retinal pigment epithelium occurs on porous polytetrafluoroethylene substrates independently of the surface chemistry

Kearns

Tasker

Zhuola

et al. 2017

J Mater Sci: Mater Med

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“…The tight junction of cells is vital to maintain epithelial morphology and restrain EMT occurrence [65]. In this study, hiPSC-RPE cells could retain their epithelial morphology and barrier function as shown by Matrigel and Activin-A (MA) treatment based on our previous study [37]. Their morphological features showed that hiPSC-RPE cells were similar to hRPE cells but not hiPSCs.…”

Section: Discussionmentioning

confidence: 79%

“…A TEER assay was used to assess the dynamic barrier function of the epithelioid cells [37]. Cells were seeded into 24-transwell inserts at 1×10 5 cells/cm 2 .…”

Section: Transepithelial Electrical Resistance (Teer) Assaymentioning

confidence: 99%

Spheroid transplantable and functional retinal pigment epithelium derived from non-colony dissociated human induced pluripotent stem cells

Guo¹,

Zhu²,

Lian³

et al. 2020

Preprint

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Background: Retinal pigment epithelium (RPE) cells derived from human induced pluripotent stem cells (hiPSCs) exhibit great promise in treating retinal degenerative diseases. Here, we would explore the feasibility of non-colony dissociated hiPSCs to differentiate into functional RPE cells (hiPSC-RPE), and offer an alternative transplantation method based on cell spheroids.Methods: hiPSC-RPE cells were identified using reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence assay, Western blotting, and flow cytometry assay. The functions of hiPSC-RPE cells in vitro and in vivo were assessed by fluorescein leakage test, transepithelial electrical resistance (TEER) assay, atomic force microscopy observation, POS phagocytosis assay, frozen tissue sections, live/dead assay, SA-β-Gal staining, and immunocytochemistry.Results: hiPSC-RPE cells positively expressed biomarkers of RPE cells but not iPSCs, such as CRALBP (97.4%), EMMPRIN (93.8%), Oct4 (2.1%), and Sox2 (2.0%). hiPSC-RPE cells displayed RPE-like characteristics including barrier function, phagocytic activity, and polarized membrane. The cells derived from hiPSC-RPE spheroids positively expressed Nestin and exhibited reduced SA-β-Gal staining. hiPSC-RPE cell spheroids could form monolayer on decellularized corneal matrixes (DCM). After one month of subretinal transplantation, hiPSC-RPE cell spheroids could survive and maintain segmental sheet growth in sodium iodate (NaIO3) induced RPE-degenerated chinchilla rabbits. Conclusion: This study suggested that non-colony dissociated hiPSCs were effectively differentiated into functional RPE cells, and hiPSC-RPE cell spheroids maintained segmental sheet growth in the subretinal of RPE degenerate chinchilla rabbits in vivo, which may lay the foundation for cell spheroid transplantation as an alternative method for RPE degenerative disease therapy in the future.

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“…Proteins were extracted from the ischemic penumbra of rat brains by radioimmunoprecipitation assay buffer (Pierce Chemical Co, Rockford, IL, USA) as previously described (Guo et al, 2016). A 40 mg sample of protein was then boiled by heating to 100°C for 10 min, fractionated by electrophoresis on a 10% sodium dodecyl sulfate-polyacrylamide gel, and transferred to a 0.45-mm polyvinylidene fluoride membrane (Bio-Rad, Hercules, CA, USA).…”

Section: Western Blotmentioning

confidence: 99%

Combined Treatment With 2-(2-Benzofu-Ranyl)-2-Imidazoline and Recombinant Tissue Plasminogen Activator Protects Blood–Brain Barrier Integrity in a Rat Model of Embolic Middle Cerebral Artery Occlusion

Zhang

et al. 2020

Front. Pharmacol.

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Recombinant tissue plasminogen activator (rt-PA) is used to treat acute ischemic stroke but is only effective if administered within 4.5 h after stroke onset. Delayed rt-PA treatment causes blood-brain barrier (BBB) disruption and hemorrhagic transformation. The compound 2-(-2-benzofuranyl)-2-imidazoline (2-BFI), a newly discovered antagonist of high-affinity postsynaptic N-methyl-D-aspartate (NMDA) receptors, has been shown to have neuroprotective effects in ischemia. Here, we investigated whether combining 2-BFI and rt-PA can ameliorate BBB disruption and prolong the therapeutic window in a rat model of embolic middle cerebral artery occlusion (eMCAO). Ischemia was induced in male Sprague Dawley rats by eMCAO, after which they were treated with 2-BFI (3 mg/kg) at 0.5 h in combination with rt-PA (10 mg/kg) at 6 or 8 h. Control rats were treated with saline or 2-BFI or rt-PA. Combined therapy with 2-BFI and rt-PA (6 h) reduced the infarct volume, denatured cell index, BBB permeability, and brain edema. This was associated with increased expression of aquaporin 4 (AQP4) and tight junction proteins (occludin and ZO-1) and downregulation of intercellular adhesion molecule 1 (ICAM-1) and matrix metalloproteinases 2 and 9 (MMP2 and MMP9). We conclude that 2-BFI protects the BBB from damage caused by delayed rt-PA treatment in ischemia. 2-BFI may therefore extend the therapeutic window up to 6 h after stroke onset in rats and may be a promising therapeutic strategy for humans. However, mechanisms to explain the effects oberved in the present study are not yet elucidated.

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Matrigel and Activin A promote cell-cell contact and anti-apoptotic activity in cultured human retinal pigment epithelium cells

Cited by 12 publications

References 47 publications

The formation of a functional retinal pigment epithelium occurs on porous polytetrafluoroethylene substrates independently of the surface chemistry

The formation of a functional retinal pigment epithelium occurs on porous polytetrafluoroethylene substrates independently of the surface chemistry

Spheroid transplantable and functional retinal pigment epithelium derived from non-colony dissociated human induced pluripotent stem cells

Combined Treatment With 2-(2-Benzofu-Ranyl)-2-Imidazoline and Recombinant Tissue Plasminogen Activator Protects Blood–Brain Barrier Integrity in a Rat Model of Embolic Middle Cerebral Artery Occlusion

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