Fabrication of cell sheets with anisotropically aligned myotubes using thermally expandable micropatterned hydrogels

Lim, Jaeman; Jun, Indong; Lee, Yu Bin; Kim, Eun Mi; Shin, Dongsuk; Jeon, Hojeong; Park, Hansoo; Shin, Heungsoo

doi:10.1007/s13233-016-4070-0

Cited by 15 publications

(7 citation statements)

References 38 publications

(40 reference statements)

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“…The partial detachment of the PD layer formed by π–π stacking or hydrophobic interactions on the PDMS and its transfer to the hydrogel may be attributed to this process . We have exploited the temperature-sensitive properties of synthetic Tetronic–tyramine-based hydrogels in previous works. ,, Consistently, the micropattern size on the hydrogels was herein increased approximately 1.3 times within 10 min when the temperature changed from 37 to 4 °C. Rapid and accurately controlled expansion is important for the generation of mechanical force and detachment of cells cultured as a large sheet or a micro-sized strip. , …”

Section: Resultsmentioning

confidence: 80%

Fabrication of Spheroids with Uniform Size by Self-Assembly of a Micro-Scaled Cell Sheet (μCS): The Effect of Cell Contraction on Spheroid Formation

Kim

Lee

Byun

et al. 2018

ACS Appl. Mater. Interfaces

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Cell spheroid culture can be an effective approach for providing an engineered microenvironment similar to an in vivo environment. Our group had recently developed a method for harvesting uniformly sized multicellular spheroids via self-assembly of micro-scaled cell sheets (μCSs) induced by the expansion of temperature-sensitive hydrogels. However, the μCS assembly process was not fully understood. In this study, we investigated the effects of cell number, pattern shape, and contractile force of cells on spheroid formation from micropatterned (width of square pattern from 100–300 μm) hydrogels. We used human dermal fibroblasts (HDFBs) as a model cell type and cultured them for 24 and 72 h. The self-assembly of μCSs cultured on square micropatterns for 72 h rapidly occurred within 4 min after reducing the temperature from 37 to 4 °C. In addition, the size distribution of spheroids was narrower with μCSs from a 72 h culture. Treatment with a ROCK1 inhibitor disrupted cytoskeletal actin fibers and the corresponding μCSs were not detached from the hydrogel. The assembly of the μCS was also affected by the micropattern shape, and the spheroid harvest efficiency was decreased to 60% when using a circular micropattern, which was explained by the stress direction on the circular versus square micropattern upon hydrogel expansion. Therefore, we confirmed that the factors controlling cell–cell interactions are important for spheroid formation using micropatterned hydrogel systems. Finally, the μCSs with dual layers of HDFBs labeled with DiD and DiO dyes resulted in the formation of spheroids with discretely localized colors within the core and shell, respectively, which suggests an outside-in assembly of detached μCSs. In consideration of these complex environmental factors, our system could be utilized in various applications as a three-dimensional culture system to fabricate cell spheroids.

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Section: Resultsmentioning

confidence: 80%

Fabrication of Spheroids with Uniform Size by Self-Assembly of a Micro-Scaled Cell Sheet (μCS): The Effect of Cell Contraction on Spheroid Formation

Kim

Lee

Byun

et al. 2018

ACS Appl. Mater. Interfaces

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“…57 Thus, this study validates that linear microgrooves can be used to align DPCs in a manner comparable to other stem cells and induce these aligned cells to lay down an aligned ECM and form a robust cell sheet. Similarly aligned cell sheets have been previously developed for use in repairing other organ systems, 29,31,35,36,39 but this is the first study to evaluate their potential effectiveness in treating peripheral nerve injuries.…”

Section: ■ Discussionmentioning

confidence: 99%

Promoting and Orienting Axon Extension Using Scaffold-Free Dental Pulp Stem Cell Sheets

Drewry

Dailey

Rothermund

et al. 2022

ACS Biomater. Sci. Eng.

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Current treatments of facial nerve injury result in poor functional outcomes due to slow and inefficient axon regeneration and aberrant reinnervation. To address these clinical challenges, bioactive scaffold-free cell sheets were engineered using neurotrophic dental pulp stem/progenitor cells (DPCs) and their aligned extracellular matrix (ECM). DPCs endogenously supply high levels of neurotrophic factors (NTFs), growth factors capable of stimulating axonal regeneration, and an aligned ECM provides guidance cues to direct axon extension. Human DPCs were grown on a substrate comprising parallel microgrooves, inducing the cells to align and deposit a linearly aligned, collagenous ECM. The resulting cell sheets were robust and could be easily removed from the underlying substrate. DPC sheets produced NTFs at levels previously shown capable of promoting axon regeneration, and, moreover, inducing DPC alignment increased the expression of select NTFs relative to unaligned controls. Furthermore, the aligned DPC sheets were able to stimulate functional neuritogenic effects in neuron-like cells in vitro. Neuronally differentiated neuroblastoma SH-SY5Y cells produced neurites that were significantly more oriented and less branched when cultured on aligned cell sheets relative to unaligned sheets. These data demonstrate that the linearly aligned DPC sheets can biomechanically support axon regeneration and improve axonal guidance which, when applied to a facial nerve injury, will result in more accurate reinnervation. The aligned DPC sheets generated here could be used in combination with commercially available nerve conduits to enhance their bioactivity or be formed into stand-alone scaffold-free nerve conduits capable of facilitating improved facial nerve recovery.

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“…Then, when the temperature was reduced from 378C to 48C, these hydrogels were disassembled and enabled the rapid harvest of cell sheets with a striated structure of ECM and myoblasts, to the desirable substrate through conformal contact with the hydrogel. Finally, the topologic micropatterns promoted the creation of aligned myotube on the harvested cell sheet [57,148].…”

Section: Hydrogels For In Vitro Engineering Skeletal Muscle Tissuesmentioning

confidence: 99%

Hydrogel biomaterials and their therapeutic potential for muscle injuries and muscular dystrophies

Lev¹,

Seliktar²

2018

J. R. Soc. Interface.

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Muscular diseases such as muscular dystrophies and muscle injuries constitute a large group of ailments that manifest as muscle weakness, atrophy or fibrosis. Although cell therapy is a promising treatment option, the delivery and retention of cells in the muscle is difficult and prevents sustained regeneration needed for adequate functional improvements. Various types of biomaterials with different physical and chemical properties have been developed to improve the delivery of cells and/or growth factors for treating muscle injuries. Hydrogels are a family of materials with distinct advantages for use as cell delivery systems in muscle injuries and ailments, including their mild processing conditions, their similarities to natural tissue extracellular matrix, and their ability to be delivered with less invasive approaches. Moreover, hydrogels can be made to completely degrade in the body, leaving behind their biological payload in a process that can enhance the therapeutic process. For these reasons, hydrogels have shown great potential as cell delivery matrices. This paper reviews a few of the hydrogel systems currently being applied together with cell therapy and/or growth factor delivery to promote the therapeutic repair of muscle injuries and muscle wasting diseases such as muscular dystrophies.

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Fabrication of cell sheets with anisotropically aligned myotubes using thermally expandable micropatterned hydrogels

Cited by 15 publications

References 38 publications

Fabrication of Spheroids with Uniform Size by Self-Assembly of a Micro-Scaled Cell Sheet (μCS): The Effect of Cell Contraction on Spheroid Formation

Fabrication of Spheroids with Uniform Size by Self-Assembly of a Micro-Scaled Cell Sheet (μCS): The Effect of Cell Contraction on Spheroid Formation

Promoting and Orienting Axon Extension Using Scaffold-Free Dental Pulp Stem Cell Sheets

Hydrogel biomaterials and their therapeutic potential for muscle injuries and muscular dystrophies

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