Journal of Pharmaceutical Investigation

The Korean Society of Pharmaceutical Sciences and Technology (한국약제학회)
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Optimization of Experimental Conditions for In vitro P-glycoprotein Assay Using LLC-GA5 Cells

  • Ahn, A-Ra (Division of Biopharmaceutics, College of Pharmacy, Kyung Hee University) ;
  • Oh, Ju-Hee (Division of Biopharmaceutics, College of Pharmacy, Kyung Hee University) ;
  • Lee, Joo-Hyun (Division of Biopharmaceutics, College of Pharmacy, Kyung Hee University) ;
  • Lee, Young-Joo (Division of Biopharmaceutics, College of Pharmacy, Kyung Hee University)
  • Received : 2010.11.02
  • Accepted : 2010.11.23
  • Published : 2010.12.20
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Abstract

Identification of compounds that function as P-glycoprotein (P-gp) substrates or inhibitors can facilitate the selection and optimization of new drug candidates. The purpose of this study is to optimize the experimental conditions for in vitro P-gp assay using LLC-GA5 cells, which is a well-known transformant cell line derived by transfecting LLC-PK1 with human MDR1. The amount of rhodamine123 transported by the LLC-GA5 and LLC-PK1 cells was evaluated under the following experimental conditions: 3 different types of transport media, colchicine pretreatment or nontreatment of the cells in the culture media, and with and without poly-L-lysine coating of the culture plates. The assay sensitivity was found to considerably differ depending on the diluents used in the transport media. P-gp-mediated transport in LLC-GA5 cells was most clearly characterized in the Hanks' balanced salt solution based transport media. The sensitivity of P-gp-mediated transport was not changed by colchicine pretreatment or poly-L-lysine coating of the culture plates.

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References

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