Scaffold-Free Three-Dimensional Cell Culture Utilizing Micromolded Nonadhesive Hydrogels

Napolitano, Anthony P.; Dean, Dylan M.; Man, Alan J.; Youssef, Jacquelyn; Ho, Don; Rago, Adam P.; Lech, Matthew P.; Morgan, Jeffrey R.

doi:10.2144/000112591

Cited by 195 publications

(174 citation statements)

References 19 publications

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“…In this study with using the 3D Petri Dish ® technique, we saw that both cell lines at different concentrations were able to form microtissue spheroids. If the distance between cells are close, cell-cell interactions will dominate due to adhesive and cohesive forces, and cells will form microtissue spheroids, and they will be able to grow in an in vivo-like microenvironment [17,29]. SH-SY5Y microtissue spheroids had more regular morphology and the spheroids at each concentration had similar shape on 1 st and 2 nd days.…”

Section: Resultsmentioning

confidence: 99%

“…Scaffold-free cultures are advantegous in providing self-assembly of cells and true physiological interactions between different types of cells without any secondary interfering material [12][13][14]. One of the scaffold-free methods is the micromolded technique utilizing nonadhesive hydrogels, cells of either one type or of more than one type spontaneously aggregate to form 3D microtissue spheroids [12][13][14][15][16][17][18][19] providing physiologically relevant conditions and a suitable platform for testing of drugs at early stage development [16] and high throughput screening processes [9]. In this context, multi-layers of cells within the microtissue spheroid forms natural barriers to drugs as intercellular tight junctions bind cells together and block or slow down the diffusion of drugs as in human tissues [19].…”

Section: Introductionmentioning

confidence: 99%

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Comparison of Microtissue Forming Capacity of Sh-Sy5y and Sk-N-as Cell Lines

Taşdemir¹,

Ürkmez²,

Dogan³

2016

deufmd

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Two-dimensional (2D) cell culture systems are important tools for basic in vitro research. However, they form a thin monolayer structure and poorly mimic the complex in vivo conditions in terms of biochemical signals, cell-cell and cell-extracellular matrix (ECM) interactions. In this study, we performed a comparative study on SH-SY5Y and SK-N-AS neuroblastoma cell lines in terms of their microtissue forming capacity. Both cell lines are commonly used to model neurodegenerative diseases in vitro. Cells were cultured using 3D Petri Dish ® technique. The cells' microtissue forming capacity was observed morphologically and microtissues' size was analized. Results indicate that microtissue forming capacity of SH-SY5Ycell line was better than that of SK-N-AS cell line. SH-SY5Y microtissues can be used as an alternative, scaffold-free in vitro 3D model for neurodegenerative diseases and neuroblastoma research. Keywords: Cell culture, Three-dimensional (3D) Cell culture, Microtissue, SH-SY5Y, SK-N-A ÖZ

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparison of Microtissue Forming Capacity of Sh-Sy5y and Sk-N-as Cell Lines

Taşdemir¹,

Ürkmez²,

Dogan³

2016

deufmd

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“…Hydrogel thickness and cross-linking density has a significant influence on cell viability, oxygen supply, and nutrient diffusion (23)(24)(25). The hydrogel thickness on the paper-based scaffold was directly governed by the alginate concentration and reaction time.…”

Section: Resultsmentioning

confidence: 99%

Hydrogel-laden paper scaffold system for origami-based tissue engineering

Kim

Lee

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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In this study, we present a method for assembling biofunctionalized paper into a multiform structured scaffold system for reliable tissue regeneration using an origami-based approach. The surface of a paper was conformally modified with a poly(styrene-comaleic anhydride) layer via initiated chemical vapor deposition followed by the immobilization of poly-L-lysine (PLL) and deposition of Ca 2+ . This procedure ensures the formation of alginate hydrogel on the paper due to Ca 2+ diffusion. Furthermore, strong adhesion of the alginate hydrogel on the paper onto the paper substrate was achieved due to an electrostatic interaction between the alginate and PLL. The developed scaffold system was versatile and allowed area-selective cell seeding. Also, the hydrogel-laden paper could be folded freely into 3D tissue-like structures using a simple origamibased method. The cylindrically constructed paper scaffold system with chondrocytes was applied into a three-ring defect trachea in rabbits. The transplanted engineered tissues replaced the native trachea without stenosis after 4 wks. As for the custom-built scaffold system, the hydrogel-laden paper system will provide a robust and facile method for the formation of tissues mimicking native tissue constructs.paper scaffolds | origami | tissue engineering | initiated chemical vapor deposition | hydrogel T he living organ changes its shape from a sheet-like arrangement with primitive cells to mature three-dimensional (3D) structures through morphogenetic processes (1-3). To date, a wide range of biomaterials have been used for the total or partial replacement of damaged organs and/or tissue structures (4-7). As the functions of the living organ are realized by periodic changes in the spatial arrangement of tissue elements, multiform scaffold systems mimicking the native tissue are desired. Moldcasting and electrospinning, among various other methods, have been introduced to fabricate diverse scaffolds (8, 9). These fabrication processes, however, possess limitations for organlike structure productions. Although recent progress in tissue engineering has focused on using 3D printer schemes, there are still limitations such as the shortage of appropriate printing materials and technical challenges related to the sensitivity of living cells (10-12).Paper-based scaffolds have been used previously for cell culture platforms (13), high-throughput biochemical assay platforms (14), and a point-of-care diagnostic system (15). As a nature-originated substrate, paper has attracted enormous research interest for applications in tissue engineering (16). Cellulose-based paper may serve as a promising material for tissue engineering as it contains macroporous structures that allow nutrient transport and oxygenation (13). In this regard, paper origami is a simple alternative approach for fabricating a multiform scaffold. Based on computeraided design (CAD) planar figures, a variety of shaped scaffolds could be designed using biofunctionalized paper.In this report, a vapor-phase method, init...

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“…17,18 Briefly, excess medium was aspirated from the wells, and a small volume of a single-cell suspension was pipetted into the main recess of each gel. …”

Section: Production Of Self-assembled Microtissuesmentioning

confidence: 99%

“…16 In addition to the spheroid geometry, we have shown that cells can self-assemble into microtissues with complex shapes, such as rods, toroids, and honeycombs. 15,[17][18][19] In this process, known as directed self-assembly, cells are seeded into a micro-mold made from a nonadhesive hydrogel that does not interfere with the cell-to-cell interactions that drive self-assembly. The mold does, however, contain nonadhesive obstacles (e.g., posts) that direct the self-organizing tissue toward specific outcomes in terms of size and shape.…”

mentioning

confidence: 99%

Micro-Mold Design Controls the 3D Morphological Evolution of Self-Assembling Multicellular Microtissues

Svoronos

Tejavibulya

Schell

et al. 2014

Tissue Engineering Part A

Self Cite

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When seeded into nonadhesive micro-molds, cells self-assemble three-dimensional (3D) multicellular microtissues via the action of cytoskeletal-mediated contraction and cell-cell adhesion. The size and shape of the tissue is a function of the cell type and the size, shape, and obstacles of the micro-mold. In this article, we used human fibroblasts to investigate some of the elements of mold design and how they can be used to guide the morphological changes that occur as a 3D tissue self-organizes. In a loop-ended dogbone mold with two nonadhesive posts, fibroblasts formed a self-constrained tissue whose tension induced morphological changes that ultimately caused the tissue to thin and rupture. Increasing the width of the dogbone's connecting rod increased the stability, whereas increasing its length decreased the stability. Mapping the rupture points showed that the balance of cell volume between the toroid and connecting rod regions of the dogbone tissue controlled the point of rupture. When cells were treated with transforming growth factor-b1, dogbones ruptured sooner due to increased cell contraction. In mold designs to form tissues with more complex shapes such as three interconnected toroids or a honeycomb, obstacle design controlled tension and tissue morphology. When the vertical posts were changed to cones, they became tension modulators that dictated when and where tension was released in a large self-organizing tissue. By understanding how elements of mold design control morphology, we can produce better models to study organogenesis, examine 3D cell mechanics, and fabricate building parts for tissue engineering.

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Scaffold-Free Three-Dimensional Cell Culture Utilizing Micromolded Nonadhesive Hydrogels

Cited by 195 publications

References 19 publications

Comparison of Microtissue Forming Capacity of Sh-Sy5y and Sk-N-as Cell Lines

Comparison of Microtissue Forming Capacity of Sh-Sy5y and Sk-N-as Cell Lines

Hydrogel-laden paper scaffold system for origami-based tissue engineering

Micro-Mold Design Controls the 3D Morphological Evolution of Self-Assembling Multicellular Microtissues

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