Characterization of Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cell Sheets Aiming for Clinical Application

Kamao, Hiroyuki; Mandai, Michiko; Okamoto, Satoshi; Sakai, Noriko; Suga, Akiko; Sugita, Sunao; Kiryu, Junichi; Takahashi, Masayo

doi:10.1016/j.stemcr.2013.12.007

Cited by 528 publications

(496 citation statements)

References 38 publications

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“…There have been studies demonstrating changes in expression of several genes and transcription factors (e.g., cathepsin D, Pax 6, calbindin, PKC-a, and Mitf) during pluripotent stem cell differentiation 34,35 ; many of these contribute to the organization of the cytoskeleton through microfilaments, intermediate filaments, and/or microtubules [36][37][38][39][40][41][42][43] . For example, upregulation of Pax6 or PKC-a during differentiation may stabilize the cytoskeleton 44,45 .…”

Section: Resultsmentioning

confidence: 99%

High-throughput physical phenotyping of cell differentiation

et al. 2017

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In this report, we present multiparameter deformability cytometry (m-DC), in which we explore a large set of parameters describing the physical phenotypes of pluripotent cells and their derivatives. m-DC utilizes microfluidic inertial focusing and hydrodynamic stretching of single cells in conjunction with high-speed video recording to realize high-throughput characterization of over 20 different cell motion and morphology-derived parameters. Parameters extracted from videos include size, deformability, deformation kinetics, and morphology. We train support vector machines that provide evidence that these additional physical measurements improve classification of induced pluripotent stem cells, mesenchymal stem cells, neural stem cells, and their derivatives compared to size and deformability alone. In addition, we utilize visual interactive stochastic neighbor embedding to visually map the high-dimensional physical phenotypic spaces occupied by these stem cells and their progeny and the pathways traversed during differentiation. This report demonstrates the potential of m-DC for improving understanding of physical differences that arise as cells differentiate and identifying cell subpopulations in a label-free manner. Ultimately, such approaches could broaden our understanding of subtle changes in cell phenotypes and their roles in human biology.

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Section: Resultsmentioning

confidence: 99%

High-throughput physical phenotyping of cell differentiation

et al. 2017

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“…Current technologies for pluripotent stem cell expansion hardly fulfil the demands of regulatory boards (Kurtz et al, 2014) and the needs of clinical use (Kamao et al, 2014), as they are associated with low cost-efficiency (Jenkins & Farid, 2015), lack stringent quality control, or do not meet good manufacturing practices (GMP) standards (Elseberg, Salzig, & Czermak, 2015). Therefore, the development of a standardized, scalable method for pluripotent stem cell expansion that is cost-effective and compatible with automation is of great interest.…”

mentioning

confidence: 99%

Scalable stirred suspension culture for the generation of billions of human induced pluripotent stem cells using single‐use bioreactors

Kwok

Ueda

Kadari

et al. 2017

J Tissue Eng Regen Med

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The production of human induced pluripotent stem cells (hiPSCs) in quantities that are relevant for cell-based therapies and cell-loaded implants through standard adherent culture is hardly achievable and lacks process scalability. A promising approach to overcoming these hurdles is the culture of hiPSCs in suspension. In this study, stirred suspension culture vessels were investigated for their suitability in the expansion of two hiPSC lines inoculated as a single cell suspension, with a free scalability between volumes of 50 and 2400 ml. The simple and robust two-step process reported here first generates hiPSC aggregates of 324 ± 71 μm diameter in 7 days in 125 ml spinner flasks (100 ml volume). These are subsequently dissociated into a single cell suspension for inoculation in 3000 ml bioreactors (1000 ml volume), finally yielding hiPSC aggregates of 198 ± 58 μm after 7 additional days. In both spinner flasks and bioreactors, hiPSCs can be cultured as aggregates for more than 40 days in suspension, maintain an undifferentiated state as confirmed by the expression of pluripotency markers TRA-1-60, TRA-1-81, SSEA-4, OCT4, and SOX2, can differentiate into cells of all three germ layers, and can be directed to differentiate into specific lineages such as cardiomyocytes. Up to a 16-fold increase in hiPSC quantity at the 100 ml volume was achieved, corresponding to a fold increase per day of 2.28; at the 1000 ml scale, an additional 10-fold increase was achieved. Taken together, 16 × 10 6 hiPSCs were expanded into 2 × 10 9 hiPSCs in 14 days for a fold increase per day of 8.93. This quantity of hiPSCs readily meets the requirements of cell-based therapies and brings their clinical potential closer to fruition.

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“…Among these, collagen is widely used as it is also the major component of Bruch's membrane. 86 Natural materials provide the advantage of directing cell behavior by closely imitating the native extracellular matrix (ECM) of the targeted tissue, although they raise concerns such as the purity of animal-derived scaffold, risk of infection, difficulties controlling consistency of the material, and mechanical instability. 87,88 In contrast, synthetic scaffolds can be custom made, including incorporation of biocompatibility, mimicking native ECM, and biodegradability.…”

Section: Scaffold Material Fabrication Technique and Propertiesmentioning

confidence: 99%

“…99 A human clinical trial in patients for treatment of AMD using iPSC-derived RPE cells started in 2013 at the RIKEN Center for Developmental Biology in Japan. 100 Previously, no immune response had been shown after transplantation of iPSC-derived RPE in monkey 86 or rodents. 101 iPSC lines were generated from AMD patients' skin cells and differentiated into RPE cells that were then seeded onto collagen gel forming a monolayer without synthetic scaffold or matrices.…”

Section: Clinical Trialsmentioning

confidence: 99%

Induced Pluripotent Stem Cell Therapies for Degenerative Disease of the Outer Retina: Disease Modeling and Cell Replacement

Foggia

Makwana

Ali³

et al. 2016

Journal of Ocular Pharmacology and Therapeutics

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Stem cell therapies are being explored as potential treatments for retinal disease. How to replace neurons in a degenerated retina presents a continued challenge for the regenerative medicine field that, if achieved, could restore sight. The major issues are: (i) the source and availability of donor cells for transplantation; (ii) the differentiation of stem cells into the required retinal cells; and (iii) the delivery, integration, functionality, and survival of new cells in the host neural network. This review considers the use of induced pluripotent stem cells (iPSC), currently under intense investigation, as a platform for cell transplantation therapy. Moreover, patientspecific iPSC are being developed for autologous cell transplantation and as a tool for modeling specific retinal diseases, testing gene therapies, and drug screening.

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Characterization of Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cell Sheets Aiming for Clinical Application

Cited by 528 publications

References 38 publications

High-throughput physical phenotyping of cell differentiation

High-throughput physical phenotyping of cell differentiation

Scalable stirred suspension culture for the generation of billions of human induced pluripotent stem cells using single‐use bioreactors

Induced Pluripotent Stem Cell Therapies for Degenerative Disease of the Outer Retina: Disease Modeling and Cell Replacement

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