scholarly journals AAV-Mediated Gene Delivery to 3D Retinal Organoids Derived from Human Induced Pluripotent Stem Cells

2020 ◽  
Vol 21 (3) ◽  
pp. 994 ◽  
Author(s):  
Marcela Garita-Hernandez ◽  
Fiona Routet ◽  
Laure Guibbal ◽  
Hanen Khabou ◽  
Lyes Toualbi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Delivery ◽  
Cell Surface ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Transduction Efficiency ◽  
Cell Surface Receptors ◽  
Rpe Cells ◽  
Surface Receptors ◽  
Induced Pluripotent

Human induced pluripotent stem cells (hiPSCs) promise a great number of future applications to investigate retinal development, pathophysiology and cell therapies for retinal degenerative diseases. Specific approaches to genetically modulate hiPSC would be valuable for all of these applications. Vectors based on adeno-associated virus (AAV) have shown the ability for gene delivery to retinal organoids derived from hiPSCs. Thus far, little work has been carried out to investigate mechanisms of AAV-mediated gene delivery and the potential advantages of engineered AAVs to genetically modify retinal organoids. In this study, we compared the early transduction efficiency of several recombinant and engineered AAVs in hiPSC-derived RPE cells and retinal organoids in relation to the availability of their cell-surface receptors and as a function of time. The genetic variant AAV2-7m8 had a superior transduction efficiency when applied at day 44 of differentiation on retinal organoids and provided long-lasting expressions for at least 4 weeks after infection without compromising cell viability. All of the capsids we tested transduced the hiPSC-RPE cells, with the AAV2-7m8 variant being the most efficient. Transduction efficiency was correlated with the presence of primary cell-surface receptors on the hiPS-derived organoids. Our study explores some of the mechanisms of cell attachment of AAVs and reports long-term gene expression resulting from gene delivery in retinal organoids.

An insight into non-integrative gene delivery approaches to generate transgene-free induced pluripotent stem cells

Gene ◽  
2019 ◽  
Vol 686 ◽  
pp. 146-159 ◽  
Author(s):  
Krishna Kumar Haridhasapavalan ◽  
Manash P. Borgohain ◽  
Chandrima Dey ◽  
Bitan Saha ◽  
Gloria Narayan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Delivery ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Induced Pluripotent ◽  
Insight Into

scholarly journals Growth Factor-free Chondrogenic Differentiation from Induced Pluripotent Stem Cells using Minicircle Vectors

2018 ◽  
Author(s):  
Yeri Alice Rim ◽  
Yoojun Nam ◽  
Ji Hyeon Ju
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Growth Factor ◽  
Gene Delivery ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Bone Morphogenetic Protein 2 ◽  
Viral Gene ◽  
Induced Pluripotent ◽  
Viral Gene Delivery

The human degenerative cartilage has low regenerative potential. Chondrocyte transplantation offers a promising strategy for cartilage treatment and regeneration. Currently chondrogenesis using human pluripotent stem cells are accomplished using human recombinant growth factors. Here, we differentiated human induced pluripotent stem cells (hiPSCs) into chondrocytes and cartilage pellet using minicircle vectors. Minicircles are used as a non-viral gene delivery system for gene therapy in various diseases. Non-viral gene delivery can produce growth factors without integrating into the host genome. Minicircle vectors containing bone morphogenetic protein 2 (BMP2) and transforming growth factor, beta 3 (TGFβ3) were successfully generated and delivered to hiPSC-derived outgrowth (OG) cells. Cell pellets generated using minicircle-transfected OG cells successfully differentiated into chondrogenic lineage. Chondrogenic pellets transfected with growth factor-encoding minicircles effectively recovered osteochondral defect in rat models. Taken together, this work shows the potential application of minicircles in cartilage regeneration using hiPSCs.

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

2020 ◽  
Author(s):  
Xiaoling Guo ◽  
Deliang Zhu ◽  
Ruiling Lian ◽  
Qiaolang Zeng ◽  
Sanjana Mathew ◽  
...  
Keyword(s):  
Stem Cells ◽  
Retinal Pigment Epithelium ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Great Promise ◽  
Rpe Cells ◽  
Cell Spheroids ◽  
Induced Pluripotent

Abstract 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.

scholarly journals In-Depth Characterisation of Retinal Pigment Epithelium (RPE) Cells Derived from Human Induced Pluripotent Stem Cells (hiPSC)

2014 ◽  
Vol 16 (3) ◽  
pp. 551-564 ◽  
Author(s):  
Caroline Brandl ◽  
Stephanie J. Zimmermann ◽  
Vladimir M. Milenkovic ◽  
Sibylle M. G. Rosendahl ◽  
Felix Grassmann ◽  
...  
Keyword(s):  
Stem Cells ◽  
Retinal Pigment Epithelium ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Rpe Cells ◽  
Induced Pluripotent

scholarly journals Gene Delivery and Expression Systems in Induced Pluripotent Stem Cells

2016 ◽  
pp. 121-133 ◽  
Author(s):  
Maolin Zhang ◽  
Kunimichi Niibe ◽  
Takeru Kondo ◽  
Yuya Kamano ◽  
Makio Saeki ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Delivery ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Expression Systems ◽  
Induced Pluripotent

scholarly journals Simple Generation of Human Induced Pluripotent Stem Cells Using Poly-β-amino Esters As the Non-viral Gene Delivery System

2011 ◽  
Vol 286 (14) ◽  
pp. 12417-12428 ◽  
Author(s):  
Núria Montserrat ◽  
Elena Garreta ◽  
Federico González ◽  
Jordán Gutiérrez ◽  
Cristina Eguizábal ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Delivery ◽  
Induced Pluripotent Stem Cells ◽  
Delivery System ◽  
Pluripotent Stem Cells ◽  
Viral Gene ◽  
Gene Delivery System ◽  
Amino Esters ◽  
Induced Pluripotent ◽  
Viral Gene Delivery
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