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Industrial Biotechnology
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2020
Cell culture is the process by which cells are grown under controlled conditions. In practice, the term “cell culture” has come to refer to the culturing of cells derived from multicellular eukaryotes, especially animal cells. The historical development and methods of cell culture are closely interrelated to those of tissue culture and organ culture. Animal cell culture became a common laboratory technique in the mid-1900s, but the concept of maintaining live cell lines separated from their original tissue source was discovered in the nineteenth century. Cells are grown and maintained at an appropriate temperature and gas mixture (typically, 37°C, 5% CO2 for mammalian cells) in a cell incubator. Culture conditions vary widely for each cell type, and variation of conditions for a particular cell type can result in different phenotypes being expressed. Aside from temperature and gas mixture, the most commonly varied factor in culture systems is the growth medium. Recipes for growth media can vary in pH, glucose concentration, growth factors, and the presence of other nutrients. The growth factors used to supplement media are often derived from animal blood, such as calf serum. One complication of these blood-derived ingredients is the potential for contamination of the culture with viruses or prions, particularly in biotechnology medical applications. Current practice is to minimize or eliminate the use of these ingredients wherever possible, but this cannot always be accomplished.
A Strategy for Regeneration of Three-Dimensional (3D) Microtissues in Microcapsules: Aerosol Atomization Technique
Published in Naznin Sultana, Sanchita Bandyopadhyay-Ghosh, Chin Fhong Soon, Tissue Engineering Strategies for Organ Regeneration, 2020
Chin Fhong Soon, Wai Yean Leong, Kian Sek Tee, Mohd Khairul Ahmad, Nafarizal Nayan
Culturing monolayers of cells in plastic dishes is routinely performed in life sciences and cell biological studies. Currently, scientific committee has begun to realize the many limitations of monolayer or two-dimensional (2D) culture model (Antoni et al. 2015, Souza et al. 2010). 2D cell model is missing accurate representation of physiological origins in terms of the proliferation, differentiation, gene and protein expression, functionality and morphology of cells (Edmondson et al. 2014). Contrarily, the three-dimensional (3D) cell culture creates extracellular matrix where cells are permitted to grow or interact with its surroundings. 3D cell culture regenerates biological relevant tissue model that restores specific cellular activities, signaling molecules and morphological structures similar to those in vivo (Kunz-Schughart et al. 2004). The cell interactions, responses and organization occurring within a 3D context demonstrated more native like and the severe limitations of 2D culture (Edmondson et al. 2014, Soon et al. 2016). 3D cell culture is part of the effort in regenerative medicine or biotechnology to recreate living and functional tissues in vitro, in which they are needed for replacement of damaged tissues (Kang et al. 2014), cancer research, application in tissue engineering (Stevens et al. 2004), pharmacological testing and stem cell research (Sugiura et al. 2005). Microencapsulation is an intensive research area to create cell and tissue model for rehabilitation of functional tissues (Zhao et al. 2017) and therapeutics purpose (da Rocha et al. 2014, Shin et al. 2013).
Industrial biotechnology
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
Cell culture is the process by which cells are grown under controlled conditions. In practice, the term “cell culture” has come to refer to the culturing of cells derived from multicellular eukaryotes, especially animal cells. The historical development and methods of cell culture are closely interrelated to those of tissue culture and organ culture. Animal cell culture became a common laboratory technique in the mid-1900s, but the concept of maintaining live cell lines separated from their original tissue source was discovered in the nineteenth century. Cells are grown and maintained at an appropriate temperature and gas mixture (typically, 37°C, 5% CO2 for mammalian cells) in a cell incubator. Culture conditions vary widely for each cell type, and variation of conditions for a particular cell type can result in different phenotypes being expressed. Aside from temperature and gas mixture, the most commonly varied factor in culture systems is the growth medium. Recipes for growth media can vary in pH, glucose concentration, growth factors, and the presence of other nutrients. The growth factors used to supplement media are often derived from animal blood, such as calf serum. One complication of these blood-derived ingredients is the potential for contamination of the culture with viruses or prions, particularly in biotechnology medical applications. Current practice is to minimize or eliminate the use of these ingredients wherever possible, but this cannot always be accomplished.
Optimization of ultraviolet/ozone (UVO3) process conditions for the preparation of gelatin coated polystyrene (PS) microcarriers
Published in Preparative Biochemistry & Biotechnology, 2022
Mohd Azmir Arifin, Maizirwan Mel, Sia Yiik Swan, Nurhusna Samsudin, Yumi Zuhanis Has-Yun Hashim, Hamzah Mohd Salleh
Animal cell culture has emerged as one of the most important tools used in life sciences today. Techniques of cell culture are essential for studying biochemical and physiological processes, and large-scale cultures of animal cells have become the preferred system for commercial production of many biological products such as recombinant proteins, monoclonal antibodies, viral vaccines, and gene therapy vectors.[1–3] While some cell types such as lymphocytes can grow in suspension, there are significant number of cell lines with industrial potentials that require attachment to solid substratum for their survival and replication.[4,5] For mass production of biologics using these ‘anchorage dependent’ cells, an economical and efficient cultivation system with extensive surface area must be established. Several systems that were examined to fulfill such requirements include spiral films, multiple plates, hollow fiber beds, and small beads.[4] Among these, microcarrier suspension culture that was first conceived by Van Wezel in 1967, appeared to be the most successful approach.[6] Microcarriers offer extremely high surface area to volume ratio that enables anchorage dependent cells to grow to high density in suspension cultures while maintaining their normal adherent mode.[7]
An automated approach for fibroblast cell confluency characterisation and sample handling using AIoT for bio-research and bio-manufacturing
Published in Cogent Engineering, 2023
Muaadh Shamhan, Ahmad Syahrin Idris, Siti Fauziah Toha, Muhammad Fauzi Daud, Izyan Mohd Idris, Hafizi Malik
Cell culture is the process of cultivating cells in an artificial environment under certain conditions until they reach a specific growth rate. It is an essential process used in the bio-manufacturing industry to produce a variety of biologics, including vaccines, therapeutic proteins, and monoclonal antibodies (Kantardjieff & Zhou, 2014). In cell culture biology, the term confluency describes the percentage of cells covering the surface area of a culture flask or petri dish. When confluency reaches a certain level, the cells are required to be separated into new cell cultures (subculture) to enable further expansion and continuous growth of the cells (Greb, 2017).
Poly (ethylene glycol) hydrogel scaffolds with multiscale porosity for culture of human adipose-derived stem cells
Published in Journal of Biomaterials Science, Polymer Edition, 2019
Haley H. Barnett, Abitha M. Heimbuck, India Pursell, Rachel A. Hegab, Benjamin J. Sawyer, Jamie J. Newman, Mary E. Caldorera-Moore
The rheology data provided insight into the physical properties of the hydrogel blends and how polymer blend and drying method can alter these properties. As expected, the shear elastic modulus decreased as the ratio of higher MW polymer increased in AD as well as LYO samples. In addition, drying method also caused a change in shear elastic modulus. All LYO samples that were investigated had a lower shear elasticity than that of their AD counterparts. The 17:3 LYO hydrogels had the lowest shear elastic modulus values while the 10:10 AD hydrogels had the highest shear elastic modulus values. Previously, the Young’s elastic modulus of the air dried then re-hydrated 10:10, 15:5, and 17:3 hydrogels blends [33,34] were within the physiologically relevant elasticity ranges for bone (50–60 kPa), cartilage (25–30 kPa) and muscle (8–10 kPa) [73,74]. In comparison the 10:10 AD hydrogel blend had a shear elastic modulus of ∼36.19 kPa. Previous researchers have estimated the brain to have a shear modulus in this range through modeling based off of cadaver experiments [75]. Other groups such as Nordez and Hug have utilized supersonic shear imaging to estimate the elastic modulus of muscle. These researchers estimate that at rest muscle tissue has an elasticity of ∼10–11 kPa and ∼21–23 kPa to ∼42–45 kPa at 3% and 7% of electromyographic activity [76]. As it is difficult to measure the elasticity of tissue in the body and there are some discrepancies in exact numbers, we chose to utilize the 10:10 hydrogel blend as it demonstrated a shear elastic modulus similar to estimates of physiologically relevant ranges. In addition, 10:10 thin LYO hydrogels had an average pore size closely resembling that of hASC diameter suggesting it as a potential scaffold for embedded cell culture and tissue engineering applications. Degradation analysis illustrates that all hydrogels, regardless of blend or drying method, retain their structural integrity and their swelling capabilities. Confirming that there is no hydrogel degradation indicates that we can reliably reproduce results when these scaffolds are utilized, which is especially important when they are used in conjunction with cells. The stability in cell culture conditions will allow us to further study them for use in tissue engineering applications.