-
Journal of Visualized Experiments : JoVE Mar 2017Monolayer cell culture does not adequately model the in vivo behavior of tissues, which involves complex cell-cell and cell-matrix interactions. Three-dimensional (3D)...
Monolayer cell culture does not adequately model the in vivo behavior of tissues, which involves complex cell-cell and cell-matrix interactions. Three-dimensional (3D) cell culture techniques are a recent innovation developed to address the shortcomings of adherent cell culture. While several techniques for generating tissue analogues in vitro have been developed, these methods are frequently complex, expensive to establish, require specialized equipment, and are generally limited by compatibility with only certain cell types. Here, we describe a rapid and flexible protocol for aggregating cells into multicellular 3D spheroids of consistent size that is compatible with growth of a variety of tumor and normal cell lines. We utilize varying concentrations of serum and methyl cellulose (MC) to promote anchorage-independent spheroid generation and prevent the formation of cell monolayers in a highly reproducible manner. Optimal conditions for individual cell lines can be achieved by adjusting MC or serum concentrations in the spheroid formation medium. The 3D spheroids generated can be collected for use in a wide range of applications, including cell signaling or gene expression studies, candidate drug screening, or in the study of cellular processes such as tumor cell invasion and migration. The protocol is also readily adapted to generate clonal spheroids from single cells, and can be adapted to assess anchorage-independent growth and anoikis-resistance. Overall, our protocol provides an easily modifiable method for generating and utilizing 3D cell spheroids in order to recapitulate the 3D microenvironment of tissues and model the in vivo growth of normal and tumor cells.
Topics: Cell Aggregation; Cell Culture Techniques; Cell Line, Tumor; Humans; Methylcellulose; Serum; Spheroids, Cellular; Time Factors
PubMed: 28448014
DOI: 10.3791/55544 -
International Journal of Biological... Jun 2022Thermal and rheological properties of methylcellulose (MC) and hydroxypropyl methylcellulose (HPMC) hydrogels with chitosan (CHI) were investigated to verify the...
Thermal and rheological properties of methylcellulose (MC) and hydroxypropyl methylcellulose (HPMC) hydrogels with chitosan (CHI) were investigated to verify the potential application of these blends as structured systems for oil transport (emulgel, oleogels, and bigels). FTIR confirmed hydrophobic interactions of cellulosic polymers with chitosan. In the temperature sweep, the thermosensitive hydrogels showed their reduced gel point compared to the original polymers. The gelation temperature was reduced from 66.9 °C for pure HPMC to 43.6 °C and 43.6 °C (MC pure polymer) to 39.3 °C when 30% CHI was added for both cases. The addition of 20 and 30% chitosan is enough to modify the extension of the gelation temperature of these polymers. These results indicate that the addition of chitosan enables MC and HPMC to form gels at lower temperatures, which could allow milder thermal conditions to be applied in processing oil carrier systems.
Topics: Chitosan; Hydrogels; Hypromellose Derivatives; Methylcellulose; Polymers; Rheology; Temperature
PubMed: 35413310
DOI: 10.1016/j.ijbiomac.2022.04.035 -
Biofabrication Sep 2021Three-dimensional (3D) bioprinting has emerged as an important tool to fabricate scaffolds with complex structures for tissue engineering and regenerative medicine...
Three-dimensional (3D) bioprinting has emerged as an important tool to fabricate scaffolds with complex structures for tissue engineering and regenerative medicine applications. For extrusion-based 3D bioprinting, the success of printing complex structures relies largely on the properties of bioink. Methylcellulose (MC) has been exploited as a potential bioink for 3D bioprinting due to its temperature-dependent rheological properties. However, MC is highly soluble and has low structural stability at room temperature, making it suboptimal for 3D bioprinting applications. In this study, we report a one-step synthesis protocol for modifying MC with norbornene (MCNB), which serves as a new bioink for 3D bioprinting. MCNB preserves the temperature-dependent reversible sol-gel transition and readily reacts with thiol-bearing linkers through light-mediated step-growth thiol-norbornene photopolymerization. Furthermore, we rendered the otherwise inert MC network bioactive through facile conjugation of integrin-binding ligands (e.g. CRGDS) or via incorporating cell-adhesive and protease-sensitive gelatin-based macromer (e.g. GelNB). The adaptability of the new MCNB-based bioink offers an attractive option for diverse 3D bioprinting applications.
Topics: Bioprinting; Hydrogels; Methylcellulose; Norbornanes; Printing, Three-Dimensional; Tissue Engineering; Tissue Scaffolds
PubMed: 34496360
DOI: 10.1088/1758-5090/ac24dc -
Biomacromolecules Jun 2022By forming and directionally freezing an aqueous foam containing cellulose nanofibrils, methylcellulose, and tannic acid, we produced a stiff and tough anisotropic solid...
By forming and directionally freezing an aqueous foam containing cellulose nanofibrils, methylcellulose, and tannic acid, we produced a stiff and tough anisotropic solid foam with low radial thermal conductivity. Along the ice-templating direction, the foam was as stiff as nanocellulose-clay composites, despite being primarily methylcellulose by mass. The foam was also stiff perpendicular to the direction of ice growth, while maintaining λ < 25 mW m K for a relative humidity (RH) up to 65% and <30 mW m K at 80% RH. This work introduces the tandem use of two practical techniques, foam formation and directional freezing, to generate a low-density anisotropic material, and this strategy could be applied to other aqueous systems where foam formation is possible.
Topics: Aerosols; Cellulose; Ice; Mesembryanthemum; Methylcellulose; Water
PubMed: 35621041
DOI: 10.1021/acs.biomac.2c00313 -
Stem Cell Reports Mar 2023Progenitor cells capable of self-renewal and differentiation in the adult human pancreas are an under-explored resource for regenerative medicine. Using...
Progenitor cells capable of self-renewal and differentiation in the adult human pancreas are an under-explored resource for regenerative medicine. Using micro-manipulation and three-dimensional colony assays we identify cells within the adult human exocrine pancreas that resemble progenitor cells. Exocrine tissues were dissociated into single cells and plated into a colony assay containing methylcellulose and 5% Matrigel. A subpopulation of ductal cells formed colonies containing differentiated ductal, acinar, and endocrine lineage cells, and expanded up to 300-fold with a ROCK inhibitor. When transplanted into diabetic mice, colonies pre-treated with a NOTCH inhibitor gave rise to insulin-expressing cells. Both colonies and primary human ducts contained cells that simultaneously express progenitor transcription factors SOX9, NKX6.1, and PDX1. In addition, in silico analysis identified progenitor-like cells within ductal clusters in a single-cell RNA sequencing dataset. Therefore, progenitor-like cells capable of self-renewal and tri-lineage differentiation either pre-exist in the adult human exocrine pancreas, or readily adapt in culture.
Topics: Humans; Adult; Mice; Animals; Methylcellulose; Diabetes Mellitus, Experimental; Pancreas; Pancreatic Ducts; Stem Cells
PubMed: 36868230
DOI: 10.1016/j.stemcr.2023.02.001 -
Journal of Biomedical Materials... May 2017Culturing cells on thermoresponsive polymers enables cells to be harvested as an intact cell sheet without disrupting the extracellular matrix or compromising cell-cell... (Comparative Study)
Comparative Study
Culturing cells on thermoresponsive polymers enables cells to be harvested as an intact cell sheet without disrupting the extracellular matrix or compromising cell-cell junctions. Previously, cell sheet fabrication methods using methylcellulose (MC) gel and PNIPAAm were independently demonstrated. In this study, MC and PNIPAAm fabrication methods are detailed and the resulting cell sheets characterized in parallel studies for direct comparison of human adipose derived stromal/stem cell (hASCs) sheet formation, cell morphology, viability, proliferation, and osteogenic potential over 21 days. A cell viability study revealed that hASCs in MC and PNIPAAm cell sheets remained viable for 21 days and proliferated until confluency. Osteogenic cell sheets exhibited upregulation of alkaline phosphatase (ALP) at day 7, as well as calcium deposition at 21 days. Additionally, expression of osteocalcin (OCN), a late-stage marker of osteogenesis, was quantified at days 14 and 21 using RT-PCR. OCN was upregulated in MC cell sheets at day 14 and PNIPAAm cell sheets at days 14 and 21. These results indicate that hASCs formed into cell sheets commit to an osteogenic lineage when cultured in osteogenic conditions. Cell sheets composed of hASCs may be used for further studies of hASC differentiation or surgical delivery of undifferentiated cells to defect sites. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1346-1354, 2017.
Topics: Acrylic Resins; Adipose Tissue; Alkaline Phosphatase; Antigens, Differentiation; Cells, Cultured; Female; Humans; Male; Methylcellulose; Osteocalcin; Osteogenesis; Stem Cells
PubMed: 28130868
DOI: 10.1002/jbm.a.36014 -
Polimery W Medycynie 2014Matrix tablets are the most popular method of oral drug administration, and polymeric materials have been used broadly in matrix formulations to modify and modulate drug... (Review)
Review
Matrix tablets are the most popular method of oral drug administration, and polymeric materials have been used broadly in matrix formulations to modify and modulate drug release rate. The main goal of the system is to extend drug release profiles to maintain a constant in vivo plasma drug concentration and a consistent pharmacological effect. Polymeric matrix tablets offer a great potential as oral controlled drug delivery systems. Cellulose derivatives, like hydroxypropyl methylcellulose (HPMC) are often used as matrix formers. However, also other types of polymers can be used for this purpose including: Kollidon SR, acrylic acid polymers such as Eudragits and Carbopols. Nevertheless, polymers of natural origin like: carragens, chitosan and alginates widely used in the food and cosmetics industry are now coming to the fore of pharmaceutical research and are used in matrix tablets technology. Modern polymers allow to obtain matrix tablets by 3D printing, which enables to develop new formulation types. In this paper, the polymers used in matrix tablets technology and examples of their applications were described.
Topics: Acrylic Resins; Alginates; Chemistry, Pharmaceutical; Delayed-Action Preparations; Drug Carriers; Hydrophobic and Hydrophilic Interactions; Hypromellose Derivatives; Methylcellulose; Polymers; Polymethacrylic Acids; Povidone; Solubility; Tablets
PubMed: 25739125
DOI: No ID Found -
The Journal of Nutrition Feb 2008Hydroxypropylmethylcellulose (HPMC) and methylcellulose (MC) are modified cellulose dietary fibers that generate viscous solutions in the gastrointestinal (GI) tract.... (Randomized Controlled Trial)
Randomized Controlled Trial
Hydroxypropylmethylcellulose (HPMC) and methylcellulose (MC) are modified cellulose dietary fibers that generate viscous solutions in the gastrointestinal (GI) tract. This study assessed the effects of high viscosity (HV) HPMC, ultra-HV (UHV) HPMC, and medium viscosity MC on postprandial glucose and insulin responses in overweight and obese men and women (n = 50). After overnight fasts, subjects consumed 5 breakfast meals containing 75 g carbohydrate, each of which contained 1 of the following: 1 g HV-HPMC, 2 g HV-HPMC, 2 g UHV-HPMC, 4 g medium-viscosity MC or control (2 g cellulose). Test sequence was randomized and double-blind, except the MC test, which was last and single-blind (46 subjects completed all 5 tests). Glucose and insulin responses were determined pre-meal and for 120 min postprandially. Median (interquartile limits) peak glucose concentration was lower (P = 0.001) after the meal containing 2.0 g UHV-HPMC (7.1, 6.3-8.2 mmol/L) compared with the control meal (7.7, 6.6-8.7 mmol/L). The control did not differ from the other conditions for peak glucose or for any of the HPMC/MC conditions for glucose incremental areas under the curves (IAUC). Peak insulin was reduced (P < 0.05) for all HPMC/MC conditions compared with control. Insulin IAUC was lower than control (P < 0.001) after meals containing 2 g HV-HPMC, 2 g UHV-HPMC, and 4 g MC. GI symptoms did not differ among treatments. These findings indicate that HV-HPMC (1 and 2 g), UHV-HPMC (2 g), and MC (4 g) consumption reduced postprandial insulin excursions consistent with delayed glucose absorption.
Topics: Blood Glucose; Cross-Over Studies; Dietary Fiber; Double-Blind Method; Female; Humans; Hypromellose Derivatives; Insulin; Male; Methylcellulose; Obesity; Postprandial Period; Time Factors
PubMed: 18203894
DOI: 10.1093/jn/138.2.292 -
Journal of Bacteriology Mar 1947
Topics: Bacteria; Cellulose; Methylcellulose
PubMed: 16561270
DOI: 10.1128/jb.53.3.257-269.1947 -
Journal of Pharmacy & Pharmaceutical... 2010Hydroxypropyl methylcellulose (HPMC) is employed for a wide variety of pharmaceutical and food preparations. Its applications as viscolizing agent (thickening agent),... (Review)
Review
Hydroxypropyl methylcellulose (HPMC) is employed for a wide variety of pharmaceutical and food preparations. Its applications as viscolizing agent (thickening agent), coating polymer, bioadhesive, in solid dispersion to enhance solubility, binder in the process of granulation and in modified release formulations have been well documented. One other notable use is in the production of capsule shells, replacing the animal derived gelatin in conventional two-piece capsules. The aim of this review is to systemically survey published literature on the HPMC use in capsule shells and resolve questions regarding their suitability as a replacement for hard gelatin capsules. Future refinements in the production and filling of HPMC capsule shells and improvement in their in vivo/in vitro dissolution would ensure their superiority over hard gelatin capsules.
Topics: Administration, Oral; Capsules; Drug Compounding; Excipients; Gelatin; Hardness; Humans; Hypromellose Derivatives; Methylcellulose; Pharmaceutical Preparations; Polymers; Solubility; Water
PubMed: 21092714
DOI: 10.18433/j3k881