Stable cell line

The key function of the vector is to introduce the cDNA under control of a strong promoter and, if a stable cell line is to be developed, also to introduce resistance to a compound that is otherwise toxic to the host cell. This facilitates selection (of only vector-bearing cells grown in special media) of the minority of cells that have... 

Zahn-Zabal M, Kobr M, Girod PA, Imhof M, Chatellard P, de Jesus M, Wurm F, Mermod N (2001) Development of stable cell lines for production or regulated expression using matrix attachment regions. J Biotechnol 87(l) 29-42... 

Generation of a Stable Cell Line Constitutively Expressing the Tet Repressor... 

Noracharttiyapot, W., Nagai, Y, Matsubara, T., Miyata, M., Shimada, M., Nagata, K. and Yamazoe, Y. (2006) Construction of several human-derived stable cell lines displaying distinct profiles of CYP3A4 induction. Drug Metab. Pharmacokinet., 21, 99-108. 

RK is cloned and expressed in SF9 cells [17,23,32] cDNA encoding RK is characterized and sequenced, 561 amino acids protein [22] RK gene expression in baculovirus-infected Sf21 cells [25,30] RK gene is cloned and expressed in Pichia pastoris GS115, COS-1 cells and HEK-293 stable cell line, best in COS-1 cells with correct posttranslational modifications [28] expression of RK and mutants in COS-7 cells [33]) [17, 22, 23, 25, 28, 30, 32, 33] <4> (cDNA encoding enzyme is cloned and expressed in COS-7 cells, sequence of the 561 amino acids protein [13]) [13]... 

Clark, K. R., Voulgaropoulou, F. and Johnson, P. R. (1996). A stable cell line carrying adenovims-inducible rep and cap genes allows for infectivity titration of adeno-associated vims vectors. Gene Ther. 3, 1124—1132. 

Generation of Stable Cell Lines Expressing Chemokine Receptors... 

Below we describe the method currently used in our lab to generate stable cell lines in HEK 293 cells. 

One of the problems encountered with stable cell lines has been the loss of receptor expression as a result of long term maintenance of cells in culture. This can usually be avoided by freezing down large numbers of early passage number cells which can be revived as and when required. 

Table 32.2 summarizes expression systems that are currently used in the biotechnology industry for generating stable cell lines. Before one makes a decision as to which expression system is preferred, it is important to evaluate all aspects including productivity and time needed for cell line development, as well as potential license fee and cost. 

TABLE 32.2 Expression Systems for Establishing Stable Cell Lines... 

The aforementioned procedure was followed using a stable cell line of K562 cells expressing human a4(34. Testing results for experimental agent racemates, (S,S), and (.R,S) diastereomers are provided in Tables 2 1, respectively. 

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