scholarly journals Evaluation of the Effects of Solvents Used in the Fabrication of Microfluidic Devices on Cell Cultures

Micromachines ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 550
Author(s):  
Xiaopeng Wen ◽  
Seiichiro Takahashi ◽  
Kenji Hatakeyama ◽  
Ken-ichiro Kamei
Keyword(s):  
Cell Cultures ◽  
Vacuum Ultraviolet ◽  
Cultured Cells ◽  
Neuroblastoma Cells ◽  
Microfluidic Devices ◽  
Promising Alternative ◽  
Residual Solvent ◽  
Microphysiological Systems ◽  
Quantitative Immunofluorescence ◽  
Solvent Bonding

Microfluidic microphysiological systems (MPSs) or “organs-on-a-chip” are a promising alternative to animal models for drug screening and toxicology tests. However, most microfluidic devices employ polydimethylsiloxane (PDMS) as the structural material; and this has several drawbacks. Cyclo-olefin polymers (COPs) are more advantageous than PDMS and other thermoplastic materials because of their low drug absorption and autofluorescence. However, most COP-based microfluidic devices are fabricated by solvent bonding of the constituent parts. Notably, the remnant solvent can affect the cultured cells. This study employed a photobonding process with vacuum ultraviolet (VUV) light to fabricate microfluidic devices without using any solvent and compared their performance with that of solvent-bonded systems (using cyclohexane, dichloromethane, or toluene as the solvent) to investigate the effects of residual solvent on cell cultures. Quantitative immunofluorescence assays indicated that the coating efficiencies of extracellular matrix proteins (e.g., Matrigel and collagen I) were lower in solvent-bonded COP devices than those in VUV-bonded devices. Furthermore, the cytotoxicity of the systems was evaluated using SH-SY5Y neuroblastoma cells, and increased apoptosis was observed in the solvent-processed devices. These results provide insights into the effects of solvents used during the fabrication of microfluidic devices and can help prevent undesirable reactions and establish good manufacturing practices.

scholarly journals Evaluation of solvents used for fabrication of microphysiological systems

2021 ◽  
Author(s):  
Xiaopeng Wen ◽  
Seiichiro Takahashi ◽  
Kenji Hatakeyama ◽  
Ken-ichiro Kamei
Keyword(s):  
Extracellular Matrix ◽  
Drug Discovery ◽  
Animal Models ◽  
Vacuum Ultraviolet ◽  
Neuroblastoma Cells ◽  
Matrix Proteins ◽  
Great Promise ◽  
Microphysiological Systems ◽  
Good Manufacturing Practices ◽  
Solvent Bonding

AbstractMicrophysiological systems (MPSs) have shown great promise for the advancement of drug discovery and toxicological tests, and as an alternative to animal models. However, although several chips and systems have been reported, some important issues are yet to be addressed, such as the use of polydimethylsiloxane (PDMS). Cyclo olefin polymers (COPs) have advantages over other thermoplastic materials, but most COP-based MPSs use solvent bonding during fabrication, which can affect any cells they are used to culture. This study uses a photobonding process with vacuum ultraviolet (UVU) to produce MPSs without the need for solvents such as cyclohexane, dichloromethane, and toluene. This is then used for comparison to investigate the effects of solvents on cell cultures. Quantitative immunofluorescent assays show that the coating efficiencies of extracellular matrix proteins, such as Matrigel and collagen I, are reduced on solvent-treated COP surfaces, compared with those prepared using VUV photobonding. Furthermore, SH-SY5Y neuroblastoma cells are used to evaluate cytotoxicity. This shows that solvent-MPSs induce apoptosis, but VUV-MPSs do not. These results provide insights into solvent bonding for MPS fabrication so that undesirable reactions can be avoided. Moreover, this work may be used to standardize MPS protocols and establish good manufacturing practices.

scholarly journals Magnetic Polymers for Magnetophoretic Separation in Microfluidic Devices

2021 ◽  
Vol 7 (7) ◽  
pp. 100
Author(s):  
Lucie Descamps ◽  
Damien Le Roy ◽  
Caterina Tomba ◽  
Anne-laure Deman
Keyword(s):  
State Of The Art ◽  
Microfluidic Devices ◽  
Large Field ◽  
Magnetic Composite ◽  
Promising Alternative ◽  
Magnetic Forces ◽  
Field Gradients ◽  
Composite Polymers ◽  
Magnetophoretic Separation ◽  
Fabrication Costs

Magnetophoresis offers many advantages for manipulating magnetic targets in microsystems. The integration of micro-flux concentrators and micro-magnets allows achieving large field gradients and therefore large reachable magnetic forces. However, the associated fabrication techniques are often complex and costly, and besides, they put specific constraints on the geometries. Magnetic composite polymers provide a promising alternative in terms of simplicity and fabrication costs, and they open new perspectives for the microstructuring, design, and integration of magnetic functions. In this review, we propose a state of the art of research works implementing magnetic polymers to trap or sort magnetic micro-beads or magnetically labeled cells in microfluidic devices.

Microwave-induced, thermally assisted solvent bonding for low-cost PMMA microfluidic devices

2009 ◽  
Vol 20 (1) ◽  
pp. 015026 ◽  
Author(s):  
Mona Rahbar ◽  
Sumanpreet Chhina ◽  
Dan Sameoto ◽  
M Parameswaran
Keyword(s):  
Low Cost ◽  
Microfluidic Devices ◽  
Solvent Bonding

scholarly journals Scrapie prion proteins accumulate in the cytoplasm of persistently infected cultured cells.

1990 ◽  
Vol 110 (6) ◽  
pp. 2117-2132 ◽  
Author(s):  
A Taraboulos ◽  
D Serban ◽  
S B Prusiner
Keyword(s):  
Cultured Cells ◽  
Prion Diseases ◽  
Neuroblastoma Cells ◽  
Brain Cell ◽  
Cellular Prion Protein ◽  
Intracellular Accumulation ◽  
Golgi Stack ◽  
Glycosyl Phosphatidylinositol ◽  
The Central Nervous System

The cellular prion protein (PrPC) is a sialoglycoprotein anchored to the external surface of cells by a glycosyl phosphatidylinositol moiety. During scrapie, an abnormal PrP isoform designated PrPSc accumulates, and much evidence argues that it is a major and necessary component of the infectious prion. Based on the resistance of native PrPSc to proteolysis and to digestion with phosphatidylinositol-specific phospholipase C as well as the enhancement of PrPSc immunoreactivity after denaturation, we devised in situ immunoassays for the detection of PrPSc in cultured cells. Using these immunoassays, we identified the sites of PrPSc accumulation in scrapie-infected cultured cells. We also used these immunoassays to isolate PrPSc-producing clones from a new hamster brain cell line (HaB) and found an excellent correlation between their PrPSc content and prion infectivity titers. In scrapie-infected HaB cells as well as in scrapie-infected mouse neuroblastoma cells, most PrPSc was found to be intracellular and most localized with ligands of the Golgi marker wheat germ agglutinin. In one scrapie-infected HaB clone, PrPSc also localized extensively with MG-160, a protein resident of the medial-Golgi stack whereas this colocalization was not observed in another subclone of these cells. Whether the sites of intracellular accumulation of PrPSc are limited to a few subcellular organelles or they are highly variable remains to be determined. If the intracellular accumulation of PrPSc is found in the cells of the central nervous system, then it might be responsible for the neuronal dysfunction and degeneration which are cardinal features of prion diseases.

scholarly journals Establishment and characterization of fibroblast cultures derived from a female common hippopotamus (Hippopotamus amphibius) skin biopsy

2020 ◽  
Author(s):  
Tao Wang ◽  
Zelong Li ◽  
Jinpu Wei ◽  
Dongmin Zheng ◽  
Chen Wang ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Cultured Cells ◽  
Population Decline ◽  
Fibroblast Cell ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Fibroblast Cultures ◽  
Hippopotamus Amphibius ◽  
The Common

AbstractThe population decline in the common hippopotamus (Hippopotamus amphibius) has necessitated the preservation of their genetic resources for species conservation and research. Of all actions, cryopreservation of fibroblast cell cultures derived from animal biopsy is considered a simple but efficient means. Nevertheless, preserving viable cell cultures of the common hippopotamus has not been achieved to our knowledge. To this end, we detailed a method to establish fibroblast cell cultures from a female common hippopotamus fetus in this study. By combining the classic tissue explant direct culture and enzymatic digestion methods, we isolated a great number of cells with typical fibroblastic morphology and high viability. Characterization of the fibroblast cultures was carried out using different techniques. In short, neither bacteria/fungi nor mycoplasma was detectable in the cell cultures throughout the study. The population doubling time was 23.9 h according to the growth curve. Karyotyping based on Giemsa staining showed that cultured cells were diploid with 36 chromosomes in all, one pair of which was sex chromosomes. Mitochondrial cytochrome C oxidase subunit I gene sequence of the cultured cells was 99.26% identical with the Hippopotamus amphibius complete mitochondrial DNA sequence registered in GenBank, confirming the cells were derived from a common hippopotamus. Flow cytometry and immunofluorescence staining results revealed that the detected cells were positive for fibroblast markers, S100A4 and Vimentin. In conclusion, we isolated and characterized a new fibroblast cell culture from a common hippopotamus skin sample and the cryopreserved cells could be useful genetic materials for the future research.

scholarly journals IMMUNIZATION OF DISSOCIATED SPLEEN CELL CULTURES FROM NORMAL MICE

1967 ◽  
Vol 126 (3) ◽  
pp. 423-442 ◽  
Author(s):  
Robert I. Mishell ◽  
Richard W. Dutton
Keyword(s):  
Spleen Cell ◽  
Cell Cultures ◽  
Culture System ◽  
Cultured Cells ◽  
Inhibitory Effect ◽  
Initial Exposure ◽  
In Vitro Immunization ◽  
In Vitro Response

A culture system for cell suspensions from mouse spleens has been described. The system provides adequate conditions for in vitro immunization on initial exposure to heterologous erythrocytes. The in vitro response closely parallels that observed in vivo with respect to size, early kinetics, antigen dose, and the inhibitory effect of passive antibody. The response of cultured cells differs in two respects from that seen in vivo. There is an increase in the ability to discriminate between different varieties of homologous erythrocytes and the in vitro response does not appear to be limited by whatever mechanisms regulate the in vivo response.

Fabrication of composite microfluidic devices for local control of oxygen tension in cell cultures

Lab on a Chip ◽  
2019 ◽  
Vol 19 (2) ◽  
pp. 306-315 ◽  
Author(s):  
Yandong Gao ◽  
Gulnaz Stybayeva ◽  
Alexander Revzin
Keyword(s):  
Oxygen Tension ◽  
Cell Cultures ◽  
Local Control ◽  
Microfluidic Chip ◽  
Gas Permeability ◽  
Microfluidic Devices

We developed a microfabrication strategy that integrated two materials with different gas permeability in a single microfluidic chip to enable local control of oxygen tension for cell cultures.

scholarly journals Assessment of Cypermethrin Residues in Tobacco by a Bioelectric Recognition Assay (BERA) Neuroblastoma Cell-Based Biosensor

Chemosensors ◽  
2019 ◽  
Vol 7 (4) ◽  
pp. 58
Author(s):  
Apostolou ◽  
Mavrikou ◽  
Denaxa ◽  
Paivana ◽  
Roussos ◽  
...  
Keyword(s):  
Matrix Effects ◽  
Neuroblastoma Cell ◽  
Neuroblastoma Cells ◽  
Specific Pattern ◽  
Promising Alternative ◽  
Biosensor System ◽  
Pyrethroid Pesticide ◽  
Actual Field ◽  
Cost Efficient ◽  
Near Future

This study presents a bioelectric cell-based biosensor for the monitoring of the pyrethroid pesticide cypermethrin, a voltage-gated sodium channel blocker, in tobacco samples. For this purpose, neuroblastoma cells were used as biorecognition elements. The potential interference by the tobacco major alkaloid nicotine on the detection of cypermethrin was also studied. In addition, fluorescence microscopy revealed a specific pattern of neuroblastoma cell calcium efflux (Ca2+) after treatment with nicotine or cypermethrin. Finally, actual field-derived tobacco extracts were used for assessing matrix effects on the biosensor’s performance. The biosensor could detect cypermethrin in concentrations up to 1.5 μg mL−1 without being influenced by the presence of nicotine and possibly other tobacco alkaloids. Though not selective for cypermethrin, the neuroblastoma-based biosensor system appears to be a promising alternative to laborious analysis methodologies for rapid, high throughput and cost-efficient screening of this pyrethroid in tobacco samples in the near future.

scholarly journals Responses of Well-Differentiated Airway Epithelial Cell Cultures from Healthy Donors and Patients with Cystic Fibrosis to Burkholderia cenocepacia Infection

2004 ◽  
Vol 72 (7) ◽  
pp. 4188-4199 ◽  
Author(s):  
Umadevi Sajjan ◽  
Shaf Keshavjee ◽  
Janet Forstner
Keyword(s):  
Cystic Fibrosis ◽  
Bacterial Infection ◽  
Cell Cultures ◽  
Cultured Cells ◽  
Burkholderia Cenocepacia ◽  
Bacterial Invasion ◽  
Mucus Layer ◽  
Complex Species ◽  
Healthy Donors ◽  
Well Differentiated

ABSTRACT Well-differentiated cultures established from airway epithelia of patients with cystic fibrosis (CF cultures) exhibited goblet cell hyperplasia, increased secretion of mucus, and higher basal levels of interleukin-8 than similarly cultured cells from healthy donors. Upon apical infection with low doses (104 to 105 CFU) of Burkholderia cenocepacia isolate BC7, the two cultures gave different responses. While normal cultures trapped the added bacteria in the mucus layer, killed and/or inhibited bacterial replication, and prevented bacterial invasion of the cells, CF cultures failed to kill and/or supported the growth of bacteria, leading to invasion of underlying epithelial cells, compromised transepithelial permeability, and cell damage. Depletion of the surface mucus layer prior to bacterial infection rendered the normal cultures susceptible to bacterial invasion, but the invading bacteria were mainly confined to vacuoles within the cells and appeared to be nonviable. In contrast, bacteria that invaded cells in CF cultures were found free in the cytoplasm surrounded by intermediate filaments and also between cells. Cultured CF airway epithelium was therefore more susceptible to infection than normal epithelium. This mimics CF tissue in vivo and illustrates differences in the way epithelia in CF patients and normal subjects handle bacterial infection. In addition, we found that the CF and normal cell cultures responded differently not only to isolate BC7 but also to isolates of other B. cepacia complex species. We therefore conclude that this cell culture model is suitable for investigation of B. cepacia complex pathogenesis in CF patients.

scholarly journals Cholesterol transporter ATP-binding cassette A1 (ABCA1) is elevated in prion disease and affects PrPC and PrPSc concentrations in cultured cells

2008 ◽  
Vol 89 (6) ◽  
pp. 1525-1532 ◽  
Author(s):  
Rajeev Kumar ◽  
Denise McClain ◽  
Rebecca Young ◽  
George A. Carlson
Keyword(s):  
Prion Disease ◽  
Cultured Cells ◽  
Prion Diseases ◽  
Neuroblastoma Cells ◽  
Direct Consequence ◽  
Brain Regions ◽  
Atp Binding ◽  
N2a Cells ◽  
Atp Binding Cassette

Prion diseases are transmissible neurodegenerative disorders of prion protein (PrP) conformation. Prion replication by conversion of benign PrPC isoforms into disease-specific PrPSc isoforms is intimately involved in prion disease pathogenesis and may be initiated in cholesterol-rich caveolae-like domains (CLD). Concentrations of the cholesterol transporter ATP-binding cassette A1 protein (ABCA1) are elevated in pre-clinical scrapie prion-infected mice and in prion-infected cells in vitro. Elevation of ABCA1 in prion-infected brain is not a direct consequence of local PrPSc accumulation, indeed levels of ABCA1 are comparable in brain regions that differ dramatically in the amount of PrPSc. Similarly, ABCA1 concentrations are identical in normal mice, transgenic mice overexpressing PrP and PrP knockout mice. In contrast, PrPC and PrPSc levels, but not Prnp mRNA, were increased by overexpression of ABCA1 in N2a neuroblastoma cells and scrapie prion-infected N2a cells (ScN2a). Conversely, RNAi-mediated knock down of Abca1 expression decreased the concentrations of PrPC in N2a cells and of PrPSc in ScN2a cells. These results suggest that ABCA1's effects on PrPC levels are post-translational and may reflect an increase in of PrPC stability, mediated either indirectly by increasing membrane cholesterol and CLD formation or by other functions of ABCA1. The increased supply of PrPC available for conversion would lead to increased PrPSc formation.

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