The application of 3D micropatterning of agarose substrate for cell culture and in situ comet assays

Mercey, Emilie; Obeïd, Patricia; Glaise, Denise; Calvo-Muñoz, Maria-Luisa; Guguen‐Guillouzo, Christiane; Fouque, Brigitte

doi:10.1016/j.biomaterials.2010.01.020

Cited by 55 publications

(39 citation statements)

References 36 publications

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“…7C). Previously, HepaRG progenitor cells have been found to undergo morphological and functional differentiation process on standard cell culture environment [40,41], on fibronectin coated 3D micropatterns [42] and in bioreactor [43]. Differentiation was most evident in NFC and EC scaffolds where secretion increased 4-8 folds from the initiation of cultures.…”

Section: Three-dimensional Cell Culture Of Hepg2 and Hepargmentioning

confidence: 98%

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Bhattacharya

Malinen

Laurén

et al. 2012

Journal of Controlled Release

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Over the recent years, various materials have been introduced as potential 3D cell culture scaffolds. These include protein extracts, peptide amphiphiles, and synthetic polymers. Hydrogel scaffolds without human or animal borne components or added bioactive components are preferred from the immunological point of view. Here we demonstrate that native nanofibrillar cellulose (NFC) hydrogels derived from the abundant plant sources provide the desired functionalities. We show 1) rheological properties that allow formation of a 3D scaffold in-situ after facile injection, 2) cellular biocompatibility without added growth factors, 3) cellular polarization, and 4) differentiation of human hepatic cell lines HepaRG and HepG2. At high shear stress, the aqueous NFC has small viscosity that supports injectability, whereas at low shear stress conditions the material is converted to an elastic gel. Due to the inherent biocompatibility without any additives, we conclude that NFC generates a feasible and sustained microenvironment for 3D cell culture for potential applications, such as drug and chemical testing, tissue engineering, and cell therapy.

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Section: Three-dimensional Cell Culture Of Hepg2 and Hepargmentioning

confidence: 98%

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Bhattacharya

Malinen

Laurén

et al. 2012

Journal of Controlled Release

Self Cite

361

290

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“…Measurements of DNA damage levels were made using an image analysis system (version 1.0, IMI comet analysis software, China). The % tail DNA served as the indicator of DNA damage levels [16,17]. The comet assay was performed blindly for technicians.…”

Section: Comet Assaymentioning

confidence: 99%

Genetic variants of H2AX gene were associated with P M 2.5 -modulated DNA damage levels in Chinese Han populations

Sun

Chu

Chen

et al. 2015

Mutation Research/Fundamental and Molecular Mechanisms of Mutag

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“…Uniformity of the pattern geometry and chemistry is critical in coculture formation because both parameters can alter the natural morphology and metabolism of the studied cells (42). Within the context of SECM measurements, the dimensions of cocultured patterns should comply with the scan range provided by the instrument to enable simultaneous analysis under identical conditions, which has proven difficult in the past (42)(43)(44)(45)(46). We responded to this challenge by partly shielding the template with a thin cover slab of PDMS to produce cocultures of HeLa and HeLa-R cells in a side-by-side format (Fig.…”

Section: Oh]mentioning

confidence: 99%

Assessment of multidrug resistance on cell coculture patterns using scanning electrochemical microscopy

Kuss

Polcari

Geißler

et al. 2013

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

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The emergence of resistance to multiple unrelated chemotherapeutic drugs impedes the treatment of several cancers. Although the involvement of ATP-binding cassette transporters has long been known, there is no in situ method capable of tracking this transporter-related resistance at the single-cell level without interfering with the cell's environment or metabolism. Here, we demonstrate that scanning electrochemical microscopy (SECM) can quantitatively and noninvasively track multidrug resistance-related protein 1-dependent multidrug resistance in patterned adenocarcinoma cervical cancer cells. Nonresistant human cancer cells and their multidrug resistant variants are arranged in a side-by-side format using a stencil-based patterning scheme, allowing for precise positioning of target cells underneath the SECM sensor. SECM measurements of the patterned cells, performed with ferrocenemethanol and [Ru(NH 3 ) 6 ] 3+ serving as electrochemical indicators, are used to establish a kinetic "map" of constant-height SECM scans, free of topography contributions. The concept underlying the work described herein may help evaluate the effectiveness of treatment administration strategies targeting reduced drug efflux.ancer cells, such as lung cancer or leukemia, acquire resistance to multiple unrelated drugs in response to treatment with chemotherapeutic agents (1, 2). Resistance impedes therapeutic effectiveness, which in turn, reduces the long-term survival rate of cancer patients (2). The emergence of multidrug resistance (MDR) involves the overexpression of transmembrane proteins P-glycoprotein (P-gp) and MDR-related protein 1 (MRP1), which both belong to the family of 5′-triphosphatebinding cassette transporters (known as ABC transporters). P-gp and MRP1 act as molecular "pumps," actively removing therapeutic agents from the cancer cells, thereby preventing the drug from inducing the desired effect on the cell nucleus or cytoplasm. MDR based on P-gp is relatively well understood and involves binding of hyaluronan to the cell surface glycoprotein CD44. The resulting up-regulation of the transcriptional cofactor p300 expression and therefore the NFkappaB-specific transcriptional up-regulation lead to the production of P-gp, and with that chemoresistance in cells (3). However, the mechanism that causes MRP1-mediated MDR remains unclear.Currently, quantification of MDR relies on immunohistochemical analyses, such as real-time PCR, focusing mostly on P-gp or other members of ABC transporters (4-9). Fluorescent MRP1-specific studies were also conducted, revealing that resistant and nonresistant cancer cells have differential intracellular content of glutathione and GST, which affect their cell death mechanism during hyperthermia (10). Microvoltammetry was also used to measure the efflux of chemotherapeutic drugs from normal and MDR cancer cells on the single-cell level, with detection limits in the nanomolar range (11). Finally, flow cytometry has routinely been used to quantify and compare expression levels and activity of dif...

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The application of 3D micropatterning of agarose substrate for cell culture and in situ comet assays

Cited by 55 publications

References 36 publications

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Genetic variants of H2AX gene were associated with P M 2.5 -modulated DNA damage levels in Chinese Han populations

Assessment of multidrug resistance on cell coculture patterns using scanning electrochemical microscopy

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