A simple microsphere‐based mold to rapidly fabricate microwell arrays for multisize 3D tumor culture

Li, Zixiu; Guo, Xiaofang; Sun, Lili; Xu, Juan; Liu, Wenming; Li, Tianbao; Wang, Jinyi

doi:10.1002/bit.27257

Cited by 10 publications

(6 citation statements)

References 50 publications

(54 reference statements)

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“…2−5 In recent years, spherical hollow features have attracted considerable attention from scientists and engineers due to their wide applications in biomedical and optical fields. Extensive efforts have therefore been devoted to the construction of spherical microcavities, including thermally reflowed microball molding, 6,7 magnetassisted microbead molding, 8,9 gas expansion molding, 10−14 and droplet molding. 15−21 Due to the geometrical contraction of the spherical microcavity's opening, the replication of spherical microcavities from microballs or microbeads inevitably suffers from the difficulty of demolding or even obtaining fractured spherical microcavities.…”

Section: ■ Introductionmentioning

confidence: 99%

Controllable Fabrication of Highly Ordered Spherical Microcavity Arrays by Replica Molding of In Situ Self-Emulsified Droplets

Han,

Zhang,

Zhang

et al. 2024

ACS Appl. Mater. Interfaces

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Ordered spherical hollow micro-and nanostructures hold great appeal in the fields of cell biology and optics. However, it is extremely challenging for standard lithography techniques to achieve spherical micro-/nanocavities. In this paper, we describe a simple, cost-effective, and scalable approach to fabricate highly ordered spherical microcavity arrays by replica molding of in situ self-emulsified droplets. The in situ self-emulsion involves a two-step process: discontinuous dewetting-induced liquid partition and interfacial tension-driven liquid spherical transformation. Subsequent replica molding of the droplets creates spherical microcavity arrays. The shapes and sizes of the microcavities can be easily modulated by varying the compositions of the droplet templates or utilizing an osmotically driven water permeation. To demonstrate the utility of this method, we employed it to create a spherical microwell array for the mass production of embryoid bodies with high viability and minimal loss. In addition, we also demonstrated the optical functions of the generated spherical microcavities by using them as microlenses. We believe that our proposed method will open exciting avenues in fields ranging from regenerative medicine and microchemistry to optical applications.

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Section: ■ Introductionmentioning

confidence: 99%

Controllable Fabrication of Highly Ordered Spherical Microcavity Arrays by Replica Molding of In Situ Self-Emulsified Droplets

Han,

Zhang,

Zhang

et al. 2024

ACS Appl. Mater. Interfaces

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“…[ 24 ] These wells, however, are difficult to fabricate, requiring micromachining, subtractive etching methods, embossing of plastics or specialized deposition methods. [ 23 , 25 , 26 , 27 , 28 ] Additive fabrication approaches are superior because they use photolithographic techniques that are simple, inexpensive, and ubiquitous. However, photolithographic fabrication of wells with controlled curvature is challenging, requiring complex mask and lens systems to modulate light intensity with micrometer resolution across the array.…”

Section: Introductionmentioning

confidence: 99%

Microbowls with Controlled Concavity for Accurate Microscale Mass Spectrometry

et al. 2022

Advanced Materials

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The properties of the substrates, including their chemistry, conductivity, and micropatterning impact sample ionization efficiency and, thus, measurement sensitivity. [8][9][10][11] For example, micrometer-scale wells are useful for segregating samples of distinct compositions so they can be separately analyzed. [12][13][14] Well arrays are also compatible with active [15,16] or passive loading techniques, [12,17] to simplify the preparation of samples for analysis. However, MALDI-MS requires samples to be dried prior to analysis. When droplets are dried on flat surfaces, they tend to distribute their analytes about the perimeter due to the coffee ring effect. [18,19] Similar processes occur in cylindrical wells, leading to precipitation along the periphery [20,21] where the signal is inhibited due to laser occlusion by the walls. The result in both cases is lowered sensitivity and increased measurement variability due to inhomogeneity of the sample spots. [18,22] Bowl-shaped wells with curved bases are advantageous because, upon drying, precipitated analytes concentrate at the center in a more uniform fashion, [23] where they are efficiently ionized. [24] These wells, however, are difficult to fabricate, requiring micromachining, subtractive etching methods, embossing of plastics or specialized deposition methods. [23,[25][26][27][28] Additive fabrication approaches are superior because they use photolithographic techniques that are simple, inexpensive, and ubiquitous. However, photolithographic fabrication of wells with controlled curvature is challenging, requiring complex mask and lens systems to modulate light intensity with micrometer resolution across the array. [29,30] Consequently, simpler but inferior methods with stamping or backfilling of sharp features with polymer are more common, even though they are tedious and provide limited control. [6,[31][32][33] A superior approach would use simple photolithographic techniques without sacrificing control over well shape and curvature.In this paper, we describe a simple method to photolithographically fabricate wells with controlled curvature in SU8. This photoresist has several properties useful for mass spectrometry, including chemical robustness, precision control of surface features, and scalable fabrication. Using the approach, we fabricate curved-bottom wells at a density of 100 000 per square centimeter on a glass slide. To demonstrate the utility of these wells, we show they yield enhanced sensitivity in microscale mass spectrometry compared to cylindrical wells. Several approaches, such as the AnchorChip [34] and the μFocus plate, [35] use surface rather than physical modification to control drying and enhance MALDI MS, but do not Patterned surfaces can enhance the sensitivity of laser desorption ionization mass spectrometry by segregating and concentrating analytes, but their fabrication can be challenging. Here, a simple method to fabricate substrates patterned with micrometer-scale wells that yield more accurate and sensitive mass spectrometry mea...

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“…Microwells can be fabricated using various microfabrication techniques, which enable the formation of spheroids of desired sizes. 11 – 13 However, most microwell-based platforms are generated on plane substrates and do not completely mimic the native nanostructure of the extracellular matrix (ECM). Meanwhile, electrospun nano/microfibers are widely used as tissue engineering scaffolds because they can recapitulate aspects of native ECM for in vitro cell cultures.…”

Section: Introductionmentioning

confidence: 99%

Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands

et al. 2022

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Three-dimensional spheroid culture enhances cell-to-cell interactions among stem cells and promotes the expression of stem cell properties; however, subsequent retrieval and delivery of these cells remain a challenge. We fabricated a thermoresponsive fiber-based microwell scaffold by combining electrospinning and hydrogel micropatterning. The resultant scaffold appeared to facilitate the formation of cellular spheroids of uniform size and enabled the expression of more stem cell-secreting growth factor genes ( EGF, IGF-1, FGF1, FGF2, and HGF), pluripotent stem cell-related genes ( SOX2 and NANOG), and adult epithelial stem cell-related genes ( LGR4, LGR5, and LGR6) than salivary gland stem cells in a monolayer culture (SGSCmonolayer). The spheroids could be retrieved efficiently by decreasing temperature. SGSC-derived spheroid (SGSCspheroid) cells were then implanted into the submandibular glands of mice at 2 weeks after fractionated X-ray irradiation at a dose of 7.5 Gy/day. At 16 weeks post-irradiation, restoration of salivary function was detected only in SGSCspheroid-implanted mice. The production of submandibular acini specific mucin increased in SGSCspheroid-implanted mice, compared with PBS control. More MIST1+ mature acinar cells were preserved in the SGSCspheroid-implanted group than in the PBS control group. Intriguingly, SGSCspheroid-implanted mice exhibited greater amelioration of tissue damage and preservation of KRT7+ terminally differentiated luminal ductal cells than SGSCmonolayer-implanted mice. The SGSCspheroid-implanted mice also showed less DNA damage and apoptotic cell death than the SGSCmonolayer-implanted mice at 2 weeks post-implantation. Additionally, a significant increase in Ki67+AQP5+ proliferative acinar cells was noted only in SGSCspheroid-implanted mice. Our results suggest that a thermoresponsive fiber-based scaffold could be of use to facilitate the production of function-enhanced SGSCspheroid cells and their subsequent retrieval and delivery to damaged salivary glands to alleviate radiation-induced apoptotic cell death and promote salivary gland regeneration.

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A simple microsphere‐based mold to rapidly fabricate microwell arrays for multisize 3D tumor culture

Cited by 10 publications

References 50 publications

Controllable Fabrication of Highly Ordered Spherical Microcavity Arrays by Replica Molding of In Situ Self-Emulsified Droplets

Controllable Fabrication of Highly Ordered Spherical Microcavity Arrays by Replica Molding of In Situ Self-Emulsified Droplets

Microbowls with Controlled Concavity for Accurate Microscale Mass Spectrometry

Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands

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