Cell culture primary cells

Stable intracellular expression of transcriptionally active ribozymes can be achieved by viral vector-mediated delivery. Currently, retroviral vectors are the most commonly used in cell culture, primary cells... 

Established cell lines, primary cell cultures of rodents, may be used. Detailed information on in vitro and in vivo assays may be obtained in reviews of SCE methods by Latt et al. (1977, 1981), Perry and Thompson (1984) and Perry et al. (1984). The in vitro methods will be briefly explored here. 

Traub O, Butterweck A, Elfgang C, Hertlein B, Balzer K, Gergs U, Hafemann B, Willecke K Immunochemical characterization of connexin31, -37, -40, -43, and -45 in cultured primary cells, transfected cell lines and murine tissues in Kanno Y, Kataoka K, Shiba Y, Shibata Y, Shimazu T (eds) Intercellular Communication through Gap Junctions. Progress in Cell Research, vol 4. Amsterdam, Elsevier, 1994, pp 343-347. 

Meunier H, Labrie F (1982) Beta-adrenergic, CRF-ergic and dopaminergic mechanisms controlling a-MSH secretion in rat pars intermedia cells in primary cell culture. Prog Neuropsychopharmacol <5 411 115. 

Slc22/Organic Ion Transporter Family. Few studies have reported the induction of Slc22a genes by PXR activators. By using rat hepatoma RL-34 cells, cultured primary rat hepatocytes and animals, it was shown that PCN induced the expression of rat hepatic Octl (Slc22al) mRNA. Induction of Octl transporter activity was further demonstrated ex vivo by increased uptake of the prototypical substrate MPP+ into hepatocytes, which were isolated from PCN-treated rats, compared to cells, which were isolated from control animals and in vivo by an increase in biliary excretion of TEA in PCN-treated rats [130]. 

A bilayer scaffold including an outer layer of an electrospun PEUU scaffold and an inner layer of porous PEUU scaffold was developed to address the cellular-ization issue [15]. The electrospun layer provided the mechanical support, while the porous scaffold layer supplied space for cell loading and infiltration. Prior to implantation, the cells including primary cells or stem cells were uniformly seeded into the porous layer using a customized vacuum device [15,40 2]. Rat vascular smooth muscle cells were seeded into the bilayer conduit and cultured in vitro for 2 days, and then this cellularized scaffold was implanted into the rat aorta model [42]. After 8 weeks, the patency of the cellularized scaffolds increased by 75% compared to 38% of the acellular scaffold. The failed scaffolds were blocked due to the intimal hyperplasia. The patent scaffolds contained a neointimal layer consisting of multiple layers of the immature contractile smooth muscle... 

Since the CM-cotton developments were designed for healthcare applications, the CM-products were sterilized with ethylene oxide (EO). The products passed the cell culture, primary skin irritation and intracutaneous irritation tests. Safety evaluation on an animal model is yet to be completed. No adverse toxicological or environmental factors are reported for cellulose ethers in general. In fact, some purified carboxymethylcelluloses, methylcelluloses, and hydroxypropylcelluloses are approved as direct food additive [8]. A significant use of CM-cellulose is also found in pharmaceuticals for skin care products such as ointments, lotions, and creams [9]. 

Cellular screening systems can be developed with primary human cells cultured in biologically relevant contexts. The outputs of these systems are focused sets of biologically relevant parameters (gene transcription, protein... 

Product extraction Effluent and waste disposal Medium preparation Seed vessel Purification Cell free supernatant Cell biomass Production bioreactor Downstream processing Medium sterilisation Primary culture Upstream processing... 

Phloretin inhibits FATP-mediated traversing of fatty acids across lipid bilayers. Phloretin is the aglycon of phlorizin and has been used to terminate the uptake of LCFAs and VLCFAs in timed in vitro uptake assays with cultured cells or in ex vivo uptake assays with isolated primary cells. 

Animal cell cultures that are initiated from cells removed directly from the animal are called primary cultures (Figure 2). Primary cultures include both explant cultures (i.e., cultures initiated from small pieces of intact tissue), as well as cultures initiated from preparations of individual or dispersed cells (obtained from intact tissue by mechanical or proteolytic dismption). Nerve fiber explant cultures in blood plasma were among the earliest types of tissue cultures (Harrison, 1907). Cells grow out from such tissue explants and form a single layer of cells completely filling the tissue culture vessel surface. Such cell cultures are called confluent monolayers. Confluent monolayers can then be treated with trypsin, so as to remove the individual cells from the culture vessel surface. The resulting cell suspension is then transferred into other culture containers, so that more viable monolayer... 

Figure 2. Initiation of primary cultures and their establishment as cell lines. Primary cultures are initiated from tissue removed from animals. Following multiple subcul-turings, cell lines in some cases are established.

A good example of the problems encountered with the use of serum with primary cultures is illustrated by the case with cultured kidney cells. The kidney epithelial cell line MDCK grows in serum-free medium supplemented with five supplements ... 

Mitochondria-associated toxicities, such as pancreatitis, are frequently demonstrated in HlV/HCV-coinfected individuals, and may significantly influence treatment options (de Mendoza and Soriano 2005). Yet, no cell culture or animal models have been developed to predict nucleoside-induced pancreatitis. Nevertheless, an association of HCV replication and mitochondrial DNA depletion in primary human lymphocytes obtained from HIV/HCV-coinfected individuals under concomitant administration of HCV and HIV medications was demonstrated by de Mendoza and coworkers (de Mendoza et al. 2007). They claimed that the use of HCV medication together with certain antiretroviral agents seemed to enhance mitochondrial damage due to a synergistic deleterious interaction between the anti-HCV and anti-HIV drugs. In contrast, an improvement in mitochondrial content with effective... 

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