Transfected cell lines

Expression studies in Xenopus oocytes or transfected cell lines originally suggested that functional GABA-activated chloride channels could be formed by receptor subunits of each class in isolation. However, much better expression occurs with two or more subunit types in combination and it is likely that most native receptors contain at least three different subunits. Co-expression of a and /I subunits results in the assembly of... 

The study of active transport mechanisms has grown substantially in recent years, with transport proteins such as P-gp, BCRP, and MRP-2 among the most studied [59]. Several types of in vitro assays to assess substrates of transporters have been established these include assays directed toward intestinal and biliary efflux [60]. Assays that measure passive and active transport are also used to assess penetration of the blood-brain barrier. In addition to the assays described above, transfected cell lines that overexpress transporters present in the blood-brain barrier are also employed [61]. 

Fig. 20.11. Substrate quality obtained by comparing basolateral-to-apical with apical-to-basolateral transport of substrates in polarized cell monolayers of MDR1-transfected cell lines [86] plotted versus (A) the log of the air/water partition coefficient, or (B) H-bond energy (arbitrary units, EUh cf. text). Units of the air/ water partition coefficient were [M ]. Compound (concentrations in Ref. [86] in brackets) were clozapine (50 nM) (1) cyclosporin A (2 tM) (2) daunorubicin (3) dexamethasone (2 tM) (4) digoxin (2 pM) (5) domperidone (2 pM) (6) etoposide (7) flunitrazepam (500 nM) (8) haloperidol (50 nM) (9) ivermectin (50 nM) (10) loperamide (2 pM) (11) morphine (2 pM) (12) ondansetron...

Tumkosit P, Kuryatov A, Luo J, Lindstrom J (2006) Beta3 subunits promote expression and nicotine-induced up-regulation of human nicotinic alpha6 nicotinic acetylcholine receptors expressed in transfected cell lines. Mol Pharmacol 70 1358-1368 Turner DM (1975) Influence of route of administration on metabolism of [14C]nicotine in four species. Xenobiotica 5 553-561... 

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. 

MVLBisG2 Efficiently Transfects MEFs, a Hard-to-Transfect Cell Line... 

Pagliaro, L. and Praestegaard, M. (2001) Transfected cell lines as tools for high throughput screening a call for standards. J. Biomolec. Screening 6, 133-136. 

The importance of ERK activation by the 5-HT1A receptor is highlighted by the observation that the receptor can stimulate proliferation of T-lymphocytes (13), pancreatic carcinoid tumor cells (52), human small cell lung carcinoma cells (53), and transfected cell lines (25,54). In contrast, endogenous lymphocyte 5-HT1A receptors have been implicated in the inhibition of mitogenesis (55). 

The values of qp can vary significantly as a function of the expression system used and of the recombinant protein product. In general, values in the range of 1 —30 pg cell-1 day-1 are reported for stably transfected cell lines. However, Chico and Jager (2000) have reported values as high as 85 pg cell-1 day-1 for the expression of a protein using the insect cell-baculovirus expression vector system. This value approaches the maximum theoretical value that is expected for animal cells, which is 100 pg cell-1 day-1 (Ozturk, 1990). 

Transfected cell line expressing receptor of interest (see Chap. 10). 

One week after the third or fourth boost, antisera may be collected and tested for immunoreactivity by ELISA using free peptide (see Subheading 3.3.1.) or by flow cytometry using receptor transfectants, cell lines or leukocytes known to express the receptor (see Subheading 3.3.2.),... 

Nicholson, A.C., Friede, S., Pearce, A., and Silverstein, R.L., 1995, Oxidized LDL binds to CD36 on human monocyte-derived macrophages and transfected cell lines Evidence implicating the lipid moiety of the lipoprotein as the binding site, Arterioscl. Tromb. Vase. Biol. 15 269-275. 

Inside-out vesicles can be perfectly used to study single efflux processes, because preloading of cells is not necessary and because of well-defined study conditions with transporter directly facing compound concentration in the incubation medium in contrast to double-transfected cell lines (3.1.2.4.L). Therefore, these studies allow establishing structure activity relationships (SAR) for one efflux transporter only. 

O Hara CM, Tang L, Taussig R, Todd RD, O Malley KL (1996) Dopamine D2L receptor couples to G alpha i2 and G alpha i3 but not G alpha il, leading to the inhibition of adenylate cyclase in transfected cell lines. 

Szolcsanyi J, Sandor Z, Petho G, Varga A, Bolcskei K, Almasi R, Riedl Z, Hajos G, Czeh G (2004) Direct evidence for activation and desensitization of the capsaicin receptor by V-oleyldopamine on TRPVl-transfected cell, line in gene deleted mice and in the rat. Neurosci Lett 361 155-158 Taguchi H, Oishi K, Sakamoto S, Shingu K (2007) Intrathecal betamethasone for cancer pain in the lower half of the body a study of its analgesic efficacy and safety. Br J Anaesth 98 385-389... 

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