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Overview of Cell Culture Processes
Published in Wei-Shou Hu, Cell Culture Bioprocess Engineering, 2020
Early cell culture explorations aspired to establish a biological platform for scientific research, but even then the potential applications of cell culture were never far from the minds of the scientists involved. Soon after primary cells became culturable, viruses were produced in primary cell culture. Cell culture quickly began to take the place of animals and embryonated chicken eggs as the production vehicle of viral vaccines. Notably, foot and mouth disease (FMD) viruses were produced in primary calf kidney cells34 and polio vaccines were produced in primary monkey kidney cells in the 1950s.35 Subsequently, continuous cell lines became the production vehicle of viruses, including BHK cells for FMD virus36 and Vero cells for polio virus.37 Human fibroblasts MRC-5 were used in human vaccine production, while interferon was produced by FS-4 cells.13 However, as we have witnessed in the past two decades, it was recombinant DNA (rDNA) technology and the use of mammalian cells for the production of therapeutic proteins that propelled cell culture to its place as a major manufacturing workhorse. Recent advances in T cell therapy offer hope that therapeutic cells will become a new class of product of cell culture processes.
Prospects of Nanotechnology in Brain Targeting Drug Delivery
Published in Bhaskar Mazumder, Subhabrata Ray, Paulami Pal, Yashwant Pathak, Nanotechnology, 2019
Srijita Chakrabarti, Probin Kr Roy, Pronobesh Chattopadhyay, Bhaskar Mazumder
The transport of organic cations has been reported in various epithelia such as the kidney, liver, intestine, CP, and placenta (Zhang et al., 1998). The functional characteristics of organic cation transport in the various epithelia have been determined by a variety of experimental techniques, such as perfused or nonperfused tissue, isolated apical and basolateral membrane vesicles, primary cell culture, and continuous cell lines. Though the tissues used in characterizing organic cation transport mechanisms have multiple carriers with overlapping driving forces and substrate selectivities, the exact mechanisms of the transport of organic cations by particular transporters are still mostly unknown (Meijer et al., 1990; Zhang et al., 1998). Moreover, it was reported that to understand the organic cation transport mechanisms, the isolation of transporters and their expression in heterologous expression systems is essential. However, the approach of isolating the transport proteins by affinity chromatography and photoaffinity labeling (Holohan et al. 1992; Kimura et al., 1995) has been hindered by a number of intrinsic problems, many of which are related to the development of specific ligands to the native transport proteins. In addition, the low abundance of transport proteins poses a major problem in their isolation and purification.
Overview of Recent Trends in Stem Cell Bioprocessing
Published in V. Sivasubramanian, Bioprocess Engineering for a Green Environment, 2018
M. Jerold, V. Sivasubramanian, K. Vasantharaj, C. Vigneshwaran
In tissue engineering, isolating cells from the donor sources and determining their functional characterization is the preliminary stage in the fabrication of the new systematic process (Kirouac and Zandstra, 2008). Figure 15.3 shows the isolation of stem cells and the formation of cell linages. In general, different cellular products are produced from various stem cells groups. The cells are isolated from various sources such as embryonic, fetal, or adult tissues. PSCs are generated using cellular reprogramming (Takahashi and Yamanaka, 2006) for prospective clinical use. In clinical therapy, primary cells are also used for various purposes. These cells are dissociated directly from parental tissue (such as kidney or liver) using mechanical or enzymatic methods, in culture medium using suitable glass or plastic containers. The primary cell culture could be of two types depending upon the kind of cells in culture. It may be either adherent cells or suspension cells. The adherent cells are called anchorage-dependent cells because they need an attachment for their growth. They are usually derived from the tissues of organs such as kidneys, where they are immobile and embedded in connective tissue. But cells that do not require any attachment for growth or do not attach to the surface of the culture vessels are called anchorage-independent cells or suspension cells. All suspension cultures are derived from cells of the blood system because these cells are also suspended in plasma in vitro, for example, lymphocytes.
Evaluation of the proinflammatory effects of contaminated bathing water
Published in Journal of Toxicology and Environmental Health, Part A, 2019
Anas A. Sattar, Wondwossen Abate, Gyorgy Fejer, Graham Bradley, Simon K. Jackson
To the best of our knowledge, the aim of the present study was first to assess the proinflammatory effects of contaminated marine bathing waters in vitro using cell culture models by measuring proinflammatory cytokines. “Contaminated” and “Clean” bathing water samples, based upon European Union water directive (2006/7/ec) indicator levels, were used as stimulants in cell culture models to investigate potential biological relevance. Previous investigators showed that the Mono Mac six cell line is a reliable model of human blood monocytes to study the biological relevance of LPS (Bamford, Ryley, and Jackson 2007; Liu et al. 2011; Zughaier, Ryley, and Jackson 1999a, 1999b). In addition, a newly described, non-transformed primary cell culture, alveolar-like murine macrophages (MPI) were found to be a reliable in vitro model to examine pathogenesis of lung inflammation and test production of proinflammatory cytokines by LPS (Fejer et al. 2013). Therefore, in this study a novel macrophage MPI cells were employed to assess the potential health impact of contaminated marine bathing water. To examine whether the possible adverse health impact of contaminated water samples is due to LPS, an inhibitor was used as previously described (Becker et al. 2005; Liu et al. 2011).
Chitosan oligosaccharide promotes osteoclast formation by stimulating the activation of MAPK and AKT signaling pathways
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Bing-Li Bai, Zhong-Jie Xie, She-Ji Weng, Zong-Yi Wu, Hang Li, Zhou-Shan Tao, Viraj Boodhun, De-Yi Yan, Zi-Jian Shen, Jia-Hao Tang, Lei Yang
For primary cell culture, BMMs were isolated from the whole bone marrow of male 6-week-old C57BL/6 mice as described previously [15]. That is to say, cells were isolated from the femoral and tibial bone marrow. Then, BMMs were cultured in α-MEM supplemented with 10% FBS, 30 ng/mL M-CSF and 1% penicillin/streptomycin in an incubator with 5% CO2 at 37 °C until they reached 90% confluence. The BMMs were subsequently seeded in a 96-well plate at a density of 8x103 cells/well in complete α-MEM supplemented with 30 ng/ml M-CSF, 50 ng/ml RANKL and different concentrations of COS (0, 5, 50 or 500 ng/ml). Culture media were replaced every 2 days until mature osteoclasts were formed. Afterward, the cells were washed twice with PBS, fixed in 4% paraformaldehyde (PFA) for 20 min, and subjected to tartrate-resistant acid phosphatase (TRAP) staining. TRAP staining is applied for identifying osteoclasts differentiation. TRAP-positive cells with > 3 nuclei were counted under a microscope.
In vitro characterization of cutaneous immunotoxicity of immortalized human keratinocytes (HaCaT) exposed to reactive and disperse textile dyes
Published in Journal of Toxicology and Environmental Health, Part A, 2018
Daniela Morais Leme, Andrea Sehr, Tamara Grummt, Jenifer Pendiuk Gonçalves, Thiago Jacomasso, Sheila Maria Brochado Winnischofer, Francine Bittencourt Potrich, Carolina Camargo de Oliveira, Edvaldo da Silva Trindade, Danielle Palma de Oliveira
Several cell-based approaches to assess key steps in initial skin inflammatory reactions were proposed and developed based upon the role of KC as first mediators of such responses (Bauch et al. 2012; Bonifas et al. 2010; Corsini and Roggen 2009; Gibbs et al. 2013; Hennen et al. 2011; Juráňová, Franková, and Ulrichová 2017; Lamberti et al. 2004; Yang et al. 2011). In order to avoid limitations produced by cost-intensive primary cell culture and donor variability, immortalized KC, particularly HaCaT cell line, a spontaneously transformed but non-tumorigenic KC cell line, have become a valid and easy-to-operate substitute for primary cultures (Bonifas et al. 2010; Magwebeba et al. 2012; Semini et al. 2014; Yang et al. 2011). Several KC-derived inflammatory mediators were examined as biomarkers of cutaneous inflammation induced by dermally applied chemicals.