Organotypic 3D Culture in Nanoscaffold Microwells Supports Salivary Gland Stem-Cell-Based Organization

Shin, Hyun Soo; Hong, Hye Jin; Koh, Won Gun; Lim, Jae Yol

doi:10.1021/acsbiomaterials.8b00894

Cited by 44 publications

(35 citation statements)

References 31 publications

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“…2C). These results are consistent with previous studies showing that salivary cells are capable of eliciting signaling responses to secretory agonists, including adenosine triphosphate (Turner et al 1998), uridine triphosphate (Baker et al 2008), and carbachol (Baker et al 2010), when plated onto different scaffolds, such as Matrigel (Baker et al 2010), nanofibers (Shin et al 2018), hyaluronic acid (Pradhan-Bhatt et al 2013), peptide modified FHs (Nam et al 2016), and poly(ethylene glycol) (Shubin et al 2017).…”

Section: Resultssupporting

confidence: 92%

Trimers Conjugated to Fibrin Hydrogels Promote Salivary Gland Function

et al. 2020

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New strategies for tissue engineering have great potential for restoring and revitalizing impaired tissues and organs, including the use of smart hydrogels that can be modified to enhance organization and functionality of the salivary glands. For instance, monomers of laminin-111 peptides chemically conjugated to fibrin hydrogel (L1pM-FH) promote cell cluster formation in vitro and salivary gland regeneration in vivo when compared with fibrin hydrogel (FH) alone; however, L1pM-FH produce only weak expression of acinar differentiation markers in vivo (e.g., aquaporin-5 and transmembrane protein 16). Since previous studies demonstrated that a greater impact can be achieved when trimeric forms were used as compared with monomeric or dimeric forms, we investigated the extent to which trimers of laminin-111 chemically conjugated to FH (L1pT-FH) can increase the expression of acinar differentiation markers and elevate saliva secretion. In vitro studies using Par-C10 acinar cells demonstrated that when compared with L1pM-FH, L1pT-FH induced similar levels of acinar-like cell clustering, polarization, lumen formation, and calcium signaling. To assess the performance of the trimeric complex in vivo, we compared the ability of L1pM-FH and L1pT-FH to increase acinar differentiation markers and restore saliva flow rate in a salivary gland wound model of C57BL/6 mice. Our results show that L1pT-FH applied to wounded mice significantly improved the expression of the acinar differentiation markers and saliva secretion when compared with the monomeric form. Together, these positive effects of L1pT-FH warrant its future testing in additional models of hyposalivation with the ultimate goal of applying this technology in humans.

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Section: Resultssupporting

confidence: 92%

Trimers Conjugated to Fibrin Hydrogels Promote Salivary Gland Function

et al. 2020

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“…In EB formation, these two features were further shown to increase the cell packing density [ 37 ], form tighter junctions, and modulate ECM networks [ 36 , 37 ]. Previous studies also established that microwell device's 3D topography and surface coating could affect cell behaviors (e.g., proliferation, differentiation) [ 17 , 41 , 79 , 80 ] and tissue morphogenesis (e.g., surface wrinkling) [ 10 , 81 ]. Furthermore, the provided deep and highly-networked (as discussed in section 3.1 ) microwells were previously found to enhance cell aggregation, nutrient exchange, and physiological functions of spheroids [ 21 , 38 ].…”

Section: Resultsmentioning

confidence: 99%

A matrigel-free method to generate matured human cerebral organoids using 3D-Printed microwell arrays

Chen

Rengarajan

Kjar

et al. 2021

Bioactive Materials

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The current methods of generating human cerebral organoids rely excessively on the use of Matrigel or other external extracellular matrices (ECM) for cell micro-environmental modulation. Matrigel embedding is problematic for long-term culture and clinical applications due to high inconsistency and other limitations. In this study, we developed a novel microwell culture platform based on 3D printing. This platform, without using Matrigel or external signaling molecules (i.e., SMAD and Wnt inhibitors), successfully generated matured human cerebral organoids with robust formation of high-level features (i.e., wrinkling/folding, lumens, neuronal layers). The formation and timing were comparable or superior to the current Matrigel methods, yet with improved consistency. The effect of microwell geometries (curvature and resolution) and coating materials (i.e., mPEG, Lipidure, BSA) was studied, showing that mPEG outperformed all other coating materials, while curved-bottom microwells outperformed flat-bottom ones. In addition, high-resolution printing outperformed low-resolution printing by creating faithful, isotropically-shaped microwells. The trend of these effects was consistent across all developmental characteristics, including EB formation efficiency and sphericity, organoid size, wrinkling index, lumen size and thickness, and neuronal layer thickness. Overall, the microwell device that was mPEG-coated, high-resolution printed, and bottom curved demonstrated the highest efficacy in promoting organoid development. This platform provided a promising strategy for generating uniform and mature human cerebral organoids as an alternative to Matrigel/ECM-embedding methods.

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“…In another case, PEG‐patterned substrates formed with plasma exposure were coated with Matrigel and used to generate human cardiac microchambers (Figure D). A similar work was performed by fabricating PEG‐based microwells via photopatterning for supporting the organization of salivary gland stem cells . These microwells coated with electrospun PCL nanofibrous scaffolds were efficient platforms for obtaining spheroids with lager diameters than the traditional floating or Matrigel culture systems.…”

Section: Hydrogels In Organoids Formationmentioning

confidence: 97%

Advances in Hydrogels in Organoids and Organs‐on‐a‐Chip

et al. 2019

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The major motivation is to better mimic human physiology and functions at multiscales from the molecular to the cellular, tissue, organ or even whole organism level. Current model systems primarily rely on the monolayer cell cultures and animal models. Simplistic monolayer cultures have their advantages, but they are often significantly different in gene expression, epigenetics, and cell function compared to native 3D tissues. [1,2] Also, they often lack cell-cell and cell-matrix interactions, [2,3] leading to the absence of tissue specific properties. Although animal models are widely used in biomedical research, they fail to faithfully predict human responses in a physiologically relevant manner due to the significant species divergences. [4] The limitations of these systems have provided an impetus for the development of alternative cell-based 3D models in vitro that better resemble the complex functionalities of living organs. Over the last several years, organoids and organ-on-a-chip (OOC), representing the major technological breakthrough, have emerged as two distinct model systems to achieve the same goal of building 3D organotypic models in vitro by bridging the gap between animal models and monolayer cultures. These engineered models are different in terms of cell source, tissue composition, architectural variability, functional features, scales, cellular fidelity, etc. Organoids, primarily evolving from developmental principles, refer to 3D multicellular tissues by self-organization of stem cells or organ-specific progenitors, which can recapitulate the intricate architectures and functionalities of in vivo organs. [5][6][7] Recent breakthroughs in organoid technology have enabled the successful generation of a variety of human organoids, such as the brain, [8,9] intestine, [10,11] liver, [12] kidney, [13,14] lung, [15] etc. These near physiological 3D organoids have garnered momentum for their potential applications in human organ development and disease modeling, drug screening and regenerative medicine. The explosion of organoids research has been catalyzed by the significant progress of stem cell biology and availability of human stem cells. In contrast, the OOC, relying on the bioengineering design principle, is a miniaturized in vitro model that can recreate the functional units of living organs on a microfluidic cell culture device using predifferentiated cells, often cell lines. [1,16] The OOC is primarily evolved from the convergence of tissue engineering and microfabricated technologies, which is characterized by the capability to simulate cellular microenvironment by precise control over fluid flow, mechanical forces and biochemical factors. [4,17] Remarkable progress has been made Significant advances in materials, microscale technology, and stem cell biology have enabled the construction of 3D tissues and organs, which will ultimately lead to more effective diagnostics and therapy. Organoids and organs-on-a-chip (OOC), evolved from developmental biology and bioengineering principles, have e...

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Organotypic 3D Culture in Nanoscaffold Microwells Supports Salivary Gland Stem-Cell-Based Organization

Cited by 44 publications

References 31 publications

Trimers Conjugated to Fibrin Hydrogels Promote Salivary Gland Function

Trimers Conjugated to Fibrin Hydrogels Promote Salivary Gland Function

A matrigel-free method to generate matured human cerebral organoids using 3D-Printed microwell arrays

Advances in Hydrogels in Organoids and Organs‐on‐a‐Chip

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