Optimizing human embryonic stem cells differentiation efficiency by screening size-tunable homogenous embryoid bodies

Moon, Sung; Ju, Jongil; Park, Soon‐Jung; Bae, Daekyeong; Chung, Hyung Min; Lee, Sang Hoon

doi:10.1016/j.biomaterials.2014.04.001

Cited by 47 publications

(42 citation statements)

References 31 publications

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“…Passaging cells onto feeders for five passages ameliorated the efficiency of differentiation, demonstrating that the effect is reversible. Previous studies in other tissues suggest that the formation of small, uniform embryoid bodies before the induction of differentiation generates more reproducible cultures (27,28). We tested this hypothesis by plating the iPSCs in AggreWell 400 plates, with each well containing 1200 low-binding microwells within it (29).…”

Section: Resultsmentioning

confidence: 98%

Human COL7A1 -corrected induced pluripotent stem cells for the treatment of recessive dystrophic epidermolysis bullosa

et al. 2014

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Patients with recessive dystrophic epidermolysis bullosa (RDEB) lack functional type VII collagen owing to mutations in the gene COL7A1 and suffer severe blistering and chronic wounds that ultimately lead to infection and development of lethal squamous cell carcinoma. The discovery of induced pluripotent stem cells (iPSCs) and the ability to edit the genome bring the possibility to provide definitive genetic therapy through corrected autologous tissues. We generated patient-derived COL7A1-corrected epithelial keratinocyte sheets for autologous grafting. We demonstrate the utility of sequential reprogramming and adenovirus-associated viral genome editing to generate corrected iPSC banks. iPSC-derived keratinocytes were produced with minimal heterogeneity, and these cells secreted wild-type type VII collagen, resulting in stratified epidermis in vitro in organotypic cultures and in vivo in mice. Sequencing of corrected cell lines before tissue formation revealed heterogeneity of cancer-predisposing mutations, allowing us to select COL7A1-corrected banks with minimal mutational burden for downstream epidermis production. Our results provide a clinical platform to use iPSCs in the treatment of debilitating genodermatoses, such as RDEB.

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Section: Resultsmentioning

confidence: 98%

Human COL7A1 -corrected induced pluripotent stem cells for the treatment of recessive dystrophic epidermolysis bullosa

et al. 2014

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“…This spatial distribution of the polymer molecules also aids in spreading and shaping individual as well as groups of cells [31–33]. Embryonic stem cells are cultured in embryoid bodies and further differentiated using conditioned culture methods for lineage specificity [34, 35]. In an attempt to identify a stem cell delivery system, murine muscle satellite cells were cultured on 3D polyglycolic acid (PGA) scaffolds fabricated from a combination of soft lithography and thermal membrane lamination.…”

Section: Influence Of the Biophysical Microenvironment On Stem Cell Rmentioning

confidence: 99%

Nanostructured scaffold as a determinant of stem cell fate

et al. 2016

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The functionality of stem cells is tightly regulated by cues from the niche, comprising both intrinsic and extrinsic cell signals. Besides chemical and growth factors, biophysical signals are important components of extrinsic signals that dictate the stem cell properties. The materials used in the fabrication of scaffolds provide the chemical cues whereas the shape of the scaffolds provides the biophysical cues. The effect of the chemical composition of the scaffolds on stem cell fate is well researched. Biophysical signals such as nanotopography, mechanical forces, stiffness of the matrix, and roughness of the biomaterial influence the fate of stem cells. However, not much is known about their role in signaling crosstalk, stem cell maintenance, and directed differentiation. Among the various techniques for scaffold design, nanotechnology has special significance. The role of nanoscale topography in scaffold design for the regulation of stem cell behavior has gained importance in regenerative medicine. Nanotechnology allows manipulation of highly advanced surfaces/scaffolds for optimal regulation of cellular behavior. Techniques such as electrospinning, soft lithography, microfluidics, carbon nanotubes, and nanostructured hydrogel are described in this review, along with their potential usage in regenerative medicine. We have also provided a brief insight into the potential signaling crosstalk that is triggered by nanomaterials that dictate a specific outcome of stem cells. This concise review compiles recent developments in nanoscale architecture and its importance in directing stem cell differentiation for prospective therapeutic applications.

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“…Several research groups have attempted to control aggregation in order to improve differentiation efficiency . According to previous reports, small aggregates were more likely to differentiate; therefore, we speculate that the addition of high concentrations of lipids possibly negatively affected iPS cell pluripotency because the aggregates were below a certain size. Therefore, optimal concentrations of KSR or lipid‐rich albumin can be identified in terms of both cell quality and quantity during mass production.…”

Section: Discussionmentioning

confidence: 78%

“…However, it is possible that aggregation triggers initial differentiation, depending on aggregate size . Several research groups have attempted to control aggregation in order to improve differentiation efficiency . According to previous reports, small aggregates were more likely to differentiate; therefore, we speculate that the addition of high concentrations of lipids possibly negatively affected iPS cell pluripotency because the aggregates were below a certain size.…”

Section: Discussionmentioning

confidence: 86%

Serum replacement with albumin‐associated lipids prevents excess aggregation and enhances growth of induced pluripotent stem cells in suspension culture

Horiguchi

Sakai

2016

Biotechnology Progress

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Suspension culture systems are currently under investigation for the mass production of pluripotent stem (PS) cells for tissue engineering; however, the control of cell aggregation in suspension culture remains challenging. Existing methods to control aggregation such as microwell culture are difficult to scale up. To address this issue, in this study a novel method that incorporates the addition of KnockOut Serum Replacement (KSR) to the PS cell culture medium was described. The method regulated cellular aggregation and significantly improved cell growth (a 2- to 10-fold increase) without any influence on pluripotency. In addition, albumin-associated lipids as the major working ingredient of KSR responsible for this inhibition of aggregation were identified. This is one of the simplest methods described to date to control aggregation and requires only chemically synthesizable reagents. Thus, this method has the potential to simplify the mass production process of PS cells and thus lower their cost. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1009-1016, 2016.

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Optimizing human embryonic stem cells differentiation efficiency by screening size-tunable homogenous embryoid bodies

Cited by 47 publications

References 31 publications

Human COL7A1 -corrected induced pluripotent stem cells for the treatment of recessive dystrophic epidermolysis bullosa

Human COL7A1 -corrected induced pluripotent stem cells for the treatment of recessive dystrophic epidermolysis bullosa

Nanostructured scaffold as a determinant of stem cell fate

Serum replacement with albumin‐associated lipids prevents excess aggregation and enhances growth of induced pluripotent stem cells in suspension culture

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